Difference between revisions of "Team:NCTU Formosa/Notebook"
| Line 9: | Line 9: | ||
a, abbr, acronym, address, big, cite, code, | a, abbr, acronym, address, big, cite, code, | ||
del, dfn, em, font, img, ins, kbd, q, s, samp, | del, dfn, em, font, img, ins, kbd, q, s, samp, | ||
| − | small, strike, strong, sub | + | small, strike, strong, sub, tt, var, |
b, u, i, center, | b, u, i, center, | ||
dl, dt, dd, ol, ul, li, | dl, dt, dd, ol, ul, li, | ||
| Line 773: | Line 773: | ||
</td> | </td> | ||
<td> | <td> | ||
| − | <B>1</B><br><br><b>PCR of</b> Transformation product: | + | <B>1</B><br><br><b>PCR of</b><br>Transformation product: |
| − | mini: Terminator[J61048] | + | <br>mini: Terminator[J61048] |
| − | ligated product: OmpA[J23101+B0034+OmpA] A<sup>r</sup> | + | <br>ligated product: OmpA[J23101+B0034+OmpA] A<sup>r</sup> |
| − | GBP[B0034+FadL-GBP+pSB1A3] A<sup>r</sup> | + | <br>GBP[B0034+FadL-GBP+pSB1A3] A<sup>r</sup> |
| − | + | <br> | |
<b>Electrophoresis of</b> | <b>Electrophoresis of</b> | ||
| − | mini: Terminator[J61048] | + | <br>mini: Terminator[J61048] |
| − | ligated product: OmpA[J23101+B0034+OmpA] A<sup>r</sup> | + | <br>ligated product: OmpA[J23101+B0034+OmpA] A<sup>r</sup> |
| Line 791: | Line 791: | ||
<td > | <td > | ||
<B>3</B><br><br><b>Ligation of</b> | <B>3</B><br><br><b>Ligation of</b> | ||
| − | GBP[J23101+B0034+FadL-GBP+pSB1A3] A<sup>r</sup> | + | <br>GBP[J23101+B0034+FadL-GBP+pSB1A3] A<sup>r</sup> |
</td> | </td> | ||
<td> | <td> | ||
<B>4</B><br><br><b>Ligation of</b> | <B>4</B><br><br><b>Ligation of</b> | ||
| − | GBP[J23101+B0034+FadL-GBP+pSB1A3] A<sup>r</sup> | + | <br>GBP[J23101+B0034+FadL-GBP+pSB1A3] A<sup>r</sup> |
| − | + | <br> | |
5/4 | 5/4 | ||
<b>Transformation off</b> ligated products: | <b>Transformation off</b> ligated products: | ||
| − | GBP[J23101+B0034+FadL-GBP+pSB1A3] A<sup>r</sup> | + | <br>GBP[J23101+B0034+FadL-GBP+pSB1A3] A<sup>r</sup> |
| − | + | <br> | |
<b>Digestion of </b> | <b>Digestion of </b> | ||
| − | RFP[E1010+J61048] (K<sup>r</sup>) (XbaI and PstI) | + | <br>RFP[E1010+J61048] (K<sup>r</sup>) (XbaI and PstI) |
| − | + | <br> | |
<b>PCR of</b> ligated products: | <b>PCR of</b> ligated products: | ||
| − | GBP[J23101+B0034+FadL-GBP+pSB1A3] A<sup>r</sup> | + | <br>GBP[J23101+B0034+FadL-GBP+pSB1A3] A<sup>r</sup> |
| − | + | <br> | |
<b>Electrophoresis of</b> PCR products: | <b>Electrophoresis of</b> PCR products: | ||
| − | GBP[J23101+B0034+FadL-GBP+pSB1A3] A<sup>r</sup> | + | <br>GBP[J23101+B0034+FadL-GBP+pSB1A3] A<sup>r</sup> |
</td> | </td> | ||
<td> | <td> | ||
<B>5</B><br><br><b>Electrophoresis of</b> | <B>5</B><br><br><b>Electrophoresis of</b> | ||
| − | digested products: | + | <br>digested products: |
| − | RFP[E1010+J61048] (K<sup>r</sup>) (XbaI and PstI) | + | <br>RFP[E1010+J61048] (K<sup>r</sup>) (XbaI and PstI) |
| − | OmpA[J23101+B0034+OmpA] A<sup>r</sup> (EcoRI and PstI) | + | <br>OmpA[J23101+B0034+OmpA] A<sup>r</sup> (EcoRI and PstI) |
| − | GBP[FadL-GBP+J61048] K<sup>r</sup> (EcoRI and PstI) | + | <br>GBP[FadL-GBP+J61048] K<sup>r</sup> (EcoRI and PstI) |
| − | + | <br> | |
<b>PCR of</b> | <b>PCR of</b> | ||
| − | OmpA[J23101+B0034+OmpA+pSB1A3] A<sup>r</sup> | + | <br>OmpA[J23101+B0034+OmpA+pSB1A3] A<sup>r</sup> |
| − | + | <br> | |
<b>Cultivation of</b> PCR product: | <b>Cultivation of</b> PCR product: | ||
| − | RFP[E1010+J61048+pSB1K3] K<sup>r</sup> | + | <br>RFP[E1010+J61048+pSB1K3] K<sup>r</sup> |
| − | GBP[J23101+B0034+FadL-GBP+pSB1A3] A<sup>r</sup> | + | <br>GBP[J23101+B0034+FadL-GBP+pSB1A3] A<sup>r</sup> |
</td> | </td> | ||
<td > | <td > | ||
<B>6</B><br><br><b>Plasmid purification of</b> | <B>6</B><br><br><b>Plasmid purification of</b> | ||
| − | RFP[E1010+J61048+pSB1K3] K<sup>r</sup> | + | <br>RFP[E1010+J61048+pSB1K3] K<sup>r</sup> |
| − | GBP[J23101+B0034+FadL-GBP+pSB1A3] A<sup>r</sup> | + | <br>GBP[J23101+B0034+FadL-GBP+pSB1A3] A<sup>r</sup> |
| − | + | <br> | |
<b>Digestion of </b> | <b>Digestion of </b> | ||
| − | GBP[J23101+B0034+FadL-GBP+pSB1A3] A<sup>r</sup> (EcoRI and PstI) | + | <br>GBP[J23101+B0034+FadL-GBP+pSB1A3] A<sup>r</sup> (EcoRI and PstI) |
| − | + | <br> | |
<b>Cultivation of</b> PCR product: | <b>Cultivation of</b> PCR product: | ||
| − | OmpA[J23101+B0034+OmpA+pSB1A3] A<sup>r</sup> | + | <br>OmpA[J23101+B0034+OmpA+pSB1A3] A<sup>r</sup> |
| − | + | <br> | |
<b>PCR of</b> backbone: | <b>PCR of</b> backbone: | ||
| − | [pSB1C3] C<sup>r</sup> | + | <br>[pSB1C3] C<sup>r</sup> |
| − | [pSB1A3] A<sup>r</sup> | + | <br>[pSB1A3] A<sup>r</sup> |
| − | [pSB1K3] K<sup>r</sup> | + | <br>[pSB1K3] K<sup>r</sup> |
| − | + | <br> | |
<b>Electrophoresis of</b> | <b>Electrophoresis of</b> | ||
| − | PCR products: OmpA[J23101+B0034+OmpA+pSB1A3] A<sup>r</sup> | + | <br>PCR products: OmpA[J23101+B0034+OmpA+pSB1A3] A<sup>r</sup> |
| − | digested products: GBP[J23101+B0034+FadL-GBP+pSB1A3] A<sup>r</sup> (EcoRI and PstI) | + | <br>digested products: GBP[J23101+B0034+FadL-GBP+pSB1A3] A<sup>r</sup> (EcoRI and PstI) |
</td> | </td> | ||
<td> | <td> | ||
<B>7</B><br><br><b>Digestion of </b> | <B>7</B><br><br><b>Digestion of </b> | ||
| − | RFP[E1010+J61048+pSB1K3] K<sup>r</sup> (EcoRI and PstI) | + | <br>RFP[E1010+J61048+pSB1K3] K<sup>r</sup> (EcoRI and PstI) |
| − | RFP[E1010+J61048+pSB1K3] K<sup>r</sup> (XbaI and PstI) | + | <br>RFP[E1010+J61048+pSB1K3] K<sup>r</sup> (XbaI and PstI) |
| − | [pSB1C3] C<sup>r</sup> (EcoRI and PstI) | + | <br>[pSB1C3] C<sup>r</sup> (EcoRI and PstI) |
| − | [pSB1A3] A<sup>r</sup> (EcoRI and PstI) | + | <br>[pSB1A3] A<sup>r</sup> (EcoRI and PstI) |
| − | [pSB1K3] K<sup>r</sup> (EcoRI and PstI) | + | <br>[pSB1K3] K<sup>r</sup> (EcoRI and PstI) |
| − | OmpA[J23101+B0034+OmpA+pSB1A3] A<sup>r</sup> (EcoRI and PstI) | + | <br>OmpA[J23101+B0034+OmpA+pSB1A3] A<sup>r</sup> (EcoRI and PstI) |
| − | + | <br> | |
<b>Plasmid purification of</b> | <b>Plasmid purification of</b> | ||
| − | OmpA[J23101+B0034+OmpA+pSB1A3] A<sup>r</sup> | + | <br>OmpA[J23101+B0034+OmpA+pSB1A3] A<sup>r</sup> |
| − | [pSB1K3] K<sup>r</sup> | + | <br>[pSB1K3] K<sup>r</sup> |
| − | + | <br> | |
<b>Electrophoresis of</b> digested products: | <b>Electrophoresis of</b> digested products: | ||
| − | RFP[E1010+J61048+pSB1K3] K<sup>r</sup> (EcoRI and PstI) | + | <br>RFP[E1010+J61048+pSB1K3] K<sup>r</sup> (EcoRI and PstI) |
| − | + | <br> | |
| − | RFP[E1010+J61048+pSB1K3] K<sup>r</sup> (XbaI and PstI) | + | <br>RFP[E1010+J61048+pSB1K3] K<sup>r</sup> (XbaI and PstI) |
| − | [pSB1C3] C<sup>r</sup> (EcoRI and PstI) | + | <br>[pSB1C3] C<sup>r</sup> (EcoRI and PstI) |
| − | [pSB1A3] A<sup>r</sup> (EcoRI and PstI) | + | <br>[pSB1A3] A<sup>r</sup> (EcoRI and PstI) |
| − | [pSB1K3] K<sup>r</sup> (EcoRI and PstI) | + | <br>[pSB1K3] K<sup>r</sup> (EcoRI and PstI) |
| − | + | <br> | |
<b>Ligation of</b> | <b>Ligation of</b> | ||
| − | RFP[B0034+E1010+J61048+pSB1C3] C<sup>r</sup> | + | <br>RFP[B0034+E1010+J61048+pSB1C3] C<sup>r</sup> |
</td> | </td> | ||
<td> | <td> | ||
<B>8</B><br><br><b>Electrophoresis of</b> digested products: | <B>8</B><br><br><b>Electrophoresis of</b> digested products: | ||
| − | OmpA[J23101+B0034+OmpA] A<sup>r</sup> (EcoRI and PstI) | + | <br>OmpA[J23101+B0034+OmpA] A<sup>r</sup> (EcoRI and PstI) |
| − | + | <br> | |
<b>Transformation off</b> ligated products: | <b>Transformation off</b> ligated products: | ||
| − | RFP[B0034+E1010+J61048+pSB1C3] C<sup>r</sup> | + | <br>RFP[B0034+E1010+J61048+pSB1C3] C<sup>r</sup> |
| − | + | <br> | |
<b>PCR of</b> | <b>PCR of</b> | ||
| − | GBP[J23101+B0034+FadL-GBP+pSB1A3] A<sup>r</sup> | + | <br>GBP[J23101+B0034+FadL-GBP+pSB1A3] A<sup>r</sup> |
| − | + | <br> | |
<b>Electrophoresis of</b> PCR products: | <b>Electrophoresis of</b> PCR products: | ||
| − | GBP[J23101+B0034+FadL-GBP+pSB1A3] A<sup>r</sup> | + | <br>GBP[J23101+B0034+FadL-GBP+pSB1A3] A<sup>r</sup> |
| − | + | <br> | |
<b>Transformation off</b> | <b>Transformation off</b> | ||
| − | GBP[J23101+B0034+FadL-GBP+pSB1A3] A<sup>r</sup> | + | <br>GBP[J23101+B0034+FadL-GBP+pSB1A3] A<sup>r</sup> |
| − | [B0030] A<sup>r</sup> | + | <br>[B0030] A<sup>r</sup> |
| − | [J23110] A<sup>r</sup> | + | <br>[J23110] A<sup>r</sup> |
</td> | </td> | ||
<td > | <td > | ||
<B>9</B><br><br><b>PCR of</b> | <B>9</B><br><br><b>PCR of</b> | ||
| − | GBP[J23101+B0034+FadL-GBP+pSB1A3] A<sup>r</sup> | + | <br>GBP[J23101+B0034+FadL-GBP+pSB1A3] A<sup>r</sup> |
| − | [B0030] A<sup>r</sup> | + | <br>[B0030] A<sup>r</sup> |
| − | [J23110] A<sup>r</sup> | + | <br>[J23110] A<sup>r</sup> |
| − | + | <br> | |
<b>Electrophoresis of</b> PCR products: | <b>Electrophoresis of</b> PCR products: | ||
| − | GBP[J23101+B0034+FadL-GBP+pSB1A3] A<sup>r</sup> | + | <br>GBP[J23101+B0034+FadL-GBP+pSB1A3] A<sup>r</sup> |
| − | [B0030] A<sup>r</sup> | + | <br>[B0030] A<sup>r</sup> |
| − | [J23110] A<sup>r</sup> | + | <br>[J23110] A<sup>r</sup> |
| − | + | <br> | |
<b>Cultivation of</b> | <b>Cultivation of</b> | ||
| − | [B0030] A<sup>r</sup> | + | <br>[B0030] A<sup>r</sup> |
| − | [J23110] A<sup>r</sup> | + | <br>[J23110] A<sup>r</sup> |
| − | + | <br> | |
<b>Ligation of</b> | <b>Ligation of</b> | ||
| − | GBP[J23101+B0034+FadL-GBP+pSB1A3] A<sup>r</sup> | + | <br>GBP[J23101+B0034+FadL-GBP+pSB1A3] A<sup>r</sup> |
</td> | </td> | ||
| Line 915: | Line 915: | ||
<td > | <td > | ||
<B>10</B><br><br><b>Electrophoresis of</b> | <B>10</B><br><br><b>Electrophoresis of</b> | ||
| − | [B0030] A<sup>r</sup> | + | <br>[B0030] A<sup>r</sup> |
| − | + | <br> | |
<b>Cultivation of</b> | <b>Cultivation of</b> | ||
| − | [B0030] A<sup>r</sup> | + | <br>[B0030] A<sup>r</sup> |
| − | + | <br> | |
| − | <b>Plasmid purification of</b> | + | <br><b>Plasmid purification of</b> |
| − | [J23110] A<sup>r</sup> | + | <br>[J23110] A<sup>r</sup> |
| − | [B0030] A<sup>r</sup> | + | <br>[B0030] A<sup>r</sup> |
| − | GBP[J23101+B0034+FadL-GBP+pSB1A3] A<sup>r</sup> | + | <br>GBP[J23101+B0034+FadL-GBP+pSB1A3] A<sup>r</sup> |
| − | + | <br> | |
| − | + | ||
<b>Digestion of </b> | <b>Digestion of </b> | ||
| − | [B0030] A<sup>r</sup> (EcoRI and SpeI) | + | <br>[B0030] A<sup>r</sup> (EcoRI and SpeI) |
| − | [J23110] A<sup>r</sup> (EcoRI and SpeI) | + | <br>[J23110] A<sup>r</sup> (EcoRI and SpeI) |
| − | GBP[J23101+B0034+FadL-GBP+pSB1A3] A<sup>r</sup> (EcoRI and PstI) | + | <br>GBP[J23101+B0034+FadL-GBP+pSB1A3] A<sup>r</sup> (EcoRI and PstI) |
| − | GBP[B0034+FadL-GBP+pSB1A3] A<sup>r</sup> (XbaI and PstI) | + | <br>GBP[B0034+FadL-GBP+pSB1A3] A<sup>r</sup> (XbaI and PstI) |
| − | + | <br> | |
<b>Electrophoresis of</b> digested products: | <b>Electrophoresis of</b> digested products: | ||
| − | [B0030] A<sup>r</sup> (EcoRI and SpeI) | + | <br>[B0030] A<sup>r</sup> (EcoRI and SpeI) |
| − | GBP[J23101+B0034+FadL-GBP+pSB1A3] A<sup>r</sup> (EcoRI and PstI) | + | <br>GBP[J23101+B0034+FadL-GBP+pSB1A3] A<sup>r</sup> (EcoRI and PstI) |
| − | GBP[B0034+FadL-GBP+pSB1A3] A<sup>r</sup> (XbaI and PstI) | + | <br>GBP[B0034+FadL-GBP+pSB1A3] A<sup>r</sup> (XbaI and PstI) |
</td> | </td> | ||
<td> | <td> | ||
<B>11</B><br><br><b>Electrophoresis of</b> digested products: | <B>11</B><br><br><b>Electrophoresis of</b> digested products: | ||
| − | [J23110] A<sup>r</sup> (EcoRI and SpeI) | + | <br>[J23110] A<sup>r</sup> (EcoRI and SpeI) |
| − | + | <br> | |
<b>Ligation of</b> | <b>Ligation of</b> | ||
| − | RFP[B0030+E1010+J61048+pSB1C3] C<sup>r</sup> | + | <br>RFP[B0030+E1010+J61048+pSB1C3] C<sup>r</sup> |
| − | GFP[B0030+E0040+J61048+pSB1K3] K<sup>r</sup> | + | <br>GFP[B0030+E0040+J61048+pSB1K3] K<sup>r</sup> |
| − | + | <br> | |
<b>Transformation off</b> | <b>Transformation off</b> | ||
| − | RFP[B0030+E1010+J61048+pSB1C3] C<sup>r</sup> | + | <br>RFP[B0030+E1010+J61048+pSB1C3] C<sup>r</sup> |
| − | GFP[B0030+E0040+J61048+pSB1K3] K<sup>r</sup> | + | <br>GFP[B0030+E0040+J61048+pSB1K3] K<sup>r</sup> |
| − | + | <br> | |
<b>PCR of</b> | <b>PCR of</b> | ||
| − | RFP[B0030+E1010+J61048+pSB1C3] C<sup>r</sup> | + | <br>RFP[B0030+E1010+J61048+pSB1C3] C<sup>r</sup> |
| − | GFP[B0030+E0040+J61048+pSB1K3] K<sup>r</sup> | + | <br>GFP[B0030+E0040+J61048+pSB1K3] K<sup>r</sup> |
</td> | </td> | ||
<td> | <td> | ||
<B>12</B><br><br><b>PCR of</b> ligated products: | <B>12</B><br><br><b>PCR of</b> ligated products: | ||
| − | RFP[B0030+E1010+J61048+pSB1C3] C<sup>r</sup> | + | <br>RFP[B0030+E1010+J61048+pSB1C3] C<sup>r</sup> |
| − | + | <br> | |
<b>Cultivation of</b> PCR products: | <b>Cultivation of</b> PCR products: | ||
| − | RFP[B0030+E1010+J61048+pSB1C3] C<sup>r</sup> | + | <br>RFP[B0030+E1010+J61048+pSB1C3] C<sup>r</sup> |
| − | + | <br> | |
<b>Plasmid purification of</b> | <b>Plasmid purification of</b> | ||
| − | RFP[B0030+E1010+J61048+pSB1C3] C<sup>r</sup> | + | <br>RFP[B0030+E1010+J61048+pSB1C3] C<sup>r</sup> |
| − | + | <br> | |
<b>Digestion of </b> | <b>Digestion of </b> | ||
| − | RFP[B0030+E1010+J61048] C<sup>r</sup> (EcoRI and SpeI) | + | <br>RFP[B0030+E1010+J61048] C<sup>r</sup> (EcoRI and SpeI) |
| − | RFP[B0030+E1010+J61048] C<sup>r</sup> (EcoRI and PstI) | + | <br>RFP[B0030+E1010+J61048] C<sup>r</sup> (EcoRI and PstI) |
| − | + | <br> | |
<b>Electrophoresis of</b> | <b>Electrophoresis of</b> | ||
| − | digested products: | + | <br>digested products: |
| − | RFP[B0030+E1010+J61048] C<sup>r</sup> (EcoRI and SpeI) | + | <br>RFP[B0030+E1010+J61048] C<sup>r</sup> (EcoRI and SpeI) |
| − | RFP[B0030+E1010+J61048] C<sup>r</sup> (EcoRI and PstI) | + | <br>RFP[B0030+E1010+J61048] C<sup>r</sup> (EcoRI and PstI) |
| − | PCR products: RFP[B0030+E1010+J61048+pSB1C3] C<sup>r</sup> | + | <br>PCR products: RFP[B0030+E1010+J61048+pSB1C3] C<sup>r</sup> |
</td> | </td> | ||
<td > | <td > | ||
<B>13</B><br><br><b>Ligation of</b> | <B>13</B><br><br><b>Ligation of</b> | ||
| − | [J23110+B0034+pSB1K3] K<sup>r</sup> | + | <br>[J23110+B0034+pSB1K3] K<sup>r</sup> |
</td> | </td> | ||
<td> | <td> | ||
<B>14</B><br><br><b>Cultivation of</b> | <B>14</B><br><br><b>Cultivation of</b> | ||
| − | OmpA[J23101+B0034+OmpA+pSB1A3] A<sup>r</sup> | + | <br>OmpA[J23101+B0034+OmpA+pSB1A3] A<sup>r</sup> |
| − | + | <br> | |
<b>Transformation off</b> | <b>Transformation off</b> | ||
| − | [J23110+B0034+pSB1K3] K<sup>r</sup> | + | <br>[J23110+B0034+pSB1K3] K<sup>r</sup> |
| − | + | <br> | |
<b>Electrophoresis of</b> digested products: | <b>Electrophoresis of</b> digested products: | ||
| − | GBP[J23101+B0034+FadL-GBP+pSB1A3] A<sup>r</sup> (EcoRI and PstI) | + | <br>GBP[J23101+B0034+FadL-GBP+pSB1A3] A<sup>r</sup> (EcoRI and PstI) |
| − | GBP[B0034+FadL-GBP+pSB1A3] A<sup>r</sup> (XbaI and PstI) | + | <br>GBP[B0034+FadL-GBP+pSB1A3] A<sup>r</sup> (XbaI and PstI) |
| − | [pSB1K3] K<sup>r</sup> (EcoRI and PstI) | + | <br>[pSB1K3] K<sup>r</sup> (EcoRI and PstI) |
</td> | </td> | ||
<td> | <td> | ||
<B>15</B><br><br><b>Digestion of </b> | <B>15</B><br><br><b>Digestion of </b> | ||
| − | GBP[B0034+FadL-GBP+pSB1A3] A<sup>r</sup> (EcoRI and PstI) | + | <br>GBP[B0034+FadL-GBP+pSB1A3] A<sup>r</sup> (EcoRI and PstI) |
| − | + | <br> | |
<b>Ligation of</b> | <b>Ligation of</b> | ||
| − | GBP[J23101+B0034+FadL-GBP+pSB1K3] K<sup>r</sup> | + | <br>GBP[J23101+B0034+FadL-GBP+pSB1K3] K<sup>r</sup> |
</td> | </td> | ||
<td > | <td > | ||
<B>16</B><br><br><b>Transformation off</b> | <B>16</B><br><br><b>Transformation off</b> | ||
| − | GBP[J23101+B0034+FadL-GBP+pSB1K3] K<sup>r</sup> | + | <br>GBP[J23101+B0034+FadL-GBP+pSB1K3] K<sup>r</sup> |
| − | + | <br> | |
<b>Electrophoresis of</b> | <b>Electrophoresis of</b> | ||
| − | GBP[B0034+FadL-GBP+pSB1A3] A<sup>r</sup> (EcoRI and PstI) | + | <br>GBP[B0034+FadL-GBP+pSB1A3] A<sup>r</sup> (EcoRI and PstI) |
| − | GBP[B0034+FadL-GBP+pSB1A3] A<sup>r</sup> (XbaI and PstI) | + | <br>GBP[B0034+FadL-GBP+pSB1A3] A<sup>r</sup> (XbaI and PstI) |
| − | + | <br> | |
<b>Ligation of</b> | <b>Ligation of</b> | ||
| − | GBP[B0034+FadL-GBP+pSB1C3] C<sup>r</sup> | + | <br>GBP[B0034+FadL-GBP+pSB1C3] C<sup>r</sup> |
| − | + | <br> | |
<b>Transformation off</b> | <b>Transformation off</b> | ||
| − | [J23110+B0034+pSB1K3] K<sup>r</sup> | + | <br>[J23110+B0034+pSB1K3] K<sup>r</sup> |
</td> | </td> | ||
| Line 1,021: | Line 1,020: | ||
<td > | <td > | ||
<B>17</B><br><br><b>Ligation of</b> | <B>17</B><br><br><b>Ligation of</b> | ||
| − | [J23110+B0034+pSB1K3] K<sup>r</sup> | + | <br>[J23110+B0034+pSB1K3] K<sup>r</sup> |
| − | GBP[J23110+B0034+FadL-GBP+pSB1K3] K<sup>r</sup> | + | <br>GBP[J23110+B0034+FadL-GBP+pSB1K3] K<sup>r</sup> |
| − | + | <br> | |
<b>PCR of</b> | <b>PCR of</b> | ||
| − | GBP[B0034+FadL-GBP+pSB1C3] C<sup>r</sup> | + | <br>GBP[B0034+FadL-GBP+pSB1C3] C<sup>r</sup> |
| − | GBP[J23110+B0034+FadL-GBP+pSB1K3] K<sup>r</sup> | + | <br>GBP[J23110+B0034+FadL-GBP+pSB1K3] K<sup>r</sup> |
| − | + | <br> | |
<b>Electrophoresis of</b> PCR products: | <b>Electrophoresis of</b> PCR products: | ||
| − | GBP[B0034+FadL-GBP+pSB1C3] C<sup>r</sup> | + | <br>GBP[B0034+FadL-GBP+pSB1C3] C<sup>r</sup> |
| − | GBP[J23110+B0034+FadL-GBP+pSB1K3] K<sup>r</sup> | + | <br>GBP[J23110+B0034+FadL-GBP+pSB1K3] K<sup>r</sup> |
</td> | </td> | ||
<td> | <td> | ||
<B>18</B><br><br><b>Cultivation of</b> | <B>18</B><br><br><b>Cultivation of</b> | ||
| − | GBP[B0034+FadL-GBP+pSB1C3] C<sup>r</sup> | + | <br>GBP[B0034+FadL-GBP+pSB1C3] C<sup>r</sup> |
| − | + | <br> | |
<b>Plasmid purification of</b> | <b>Plasmid purification of</b> | ||
| − | GBP[B0034+FadL-GBP+pSB1C3] C<sup>r</sup> | + | <br>GBP[B0034+FadL-GBP+pSB1C3] C<sup>r</sup> |
| − | + | <br> | |
<b>Digestion of </b> | <b>Digestion of </b> | ||
| − | GBP[B0034+FadL-GBP] C<sup>r</sup> (XbaI and PstI) | + | <br>GBP[B0034+FadL-GBP] C<sup>r</sup> (XbaI and PstI) |
| − | RFP[B0030+E1010+J61048] C<sup>r</sup> (XbaI and PstI) | + | <br>RFP[B0030+E1010+J61048] C<sup>r</sup> (XbaI and PstI) |
| − | GFP[B0030+E0040+J61048] | + | <br>GFP[B0030+E0040+J61048] |
<b>Transformation off</b> | <b>Transformation off</b> | ||
| − | [J23110+B0034+pSB1K3] K<sup>r</sup> | + | <br>[J23110+B0034+pSB1K3] K<sup>r</sup> |
| − | GBP[J23101+B0034+FadL-GBP+pSB1K3] K<sup>r</sup> | + | <br>GBP[J23101+B0034+FadL-GBP+pSB1K3] K<sup>r</sup> |
| − | GFP[B0030+E0040+J61048] | + | <br>GFP[B0030+E0040+J61048] |
</td> | </td> | ||
<td> | <td> | ||
<B>19</B><br><br><b>Ligation of</b> | <B>19</B><br><br><b>Ligation of</b> | ||
| − | GBP[J23101+B0034+FadL-GBP+pSB1K3] K<sup>r</sup> | + | <br>GBP[J23101+B0034+FadL-GBP+pSB1K3] K<sup>r</sup> |
| − | + | <br> | |
<b>PCR of</b> | <b>PCR of</b> | ||
| − | GBP[J23101+B0034+FadL-GBP+pSB1K3] K<sup>r</sup> | + | <br>GBP[J23101+B0034+FadL-GBP+pSB1K3] K<sup>r</sup> |
| − | + | <br> | |
<b>Electrophoresis of</b> | <b>Electrophoresis of</b> | ||
| − | digested products: | + | <br>digested products: |
| − | RFP[B0030+E1010+J61048] C<sup>r</sup> (XbaI and PstI) | + | <br>RFP[B0030+E1010+J61048] C<sup>r</sup> (XbaI and PstI) |
| − | GBP[B0034+FadL-GBP] C<sup>r</sup> (XbaI and PstI) | + | <br>GBP[B0034+FadL-GBP] C<sup>r</sup> (XbaI and PstI) |
| − | PCR products: GBP[J23101+B0034+FadL-GBP+pSB1K3] K<sup>r</sup> | + | <br>PCR products: GBP[J23101+B0034+FadL-GBP+pSB1K3] K<sup>r</sup> |
| − | + | <br> | |
<b>Transformation off</b> | <b>Transformation off</b> | ||
| − | scFv-B[scFv-B+pSB1K3] K<sup>r</sup> | + | <br>scFv-B[scFv-B+pSB1K3] K<sup>r</sup> |
| − | scFv-C[scFv-C+pSB1K3] K<sup>r</sup> | + | <br>scFv-C[scFv-C+pSB1K3] K<sup>r</sup> |
| − | GFP[B0030+E0040+J61048] | + | <br>GFP[B0030+E0040+J61048] |
| − | + | <br> | |
<b>Cultivation of</b> | <b>Cultivation of</b> | ||
| − | GBP[J23101+B0034+FadL-GBP+pSB1K3] K<sup>r</sup> | + | <br>GBP[J23101+B0034+FadL-GBP+pSB1K3] K<sup>r</sup> |
</td> | </td> | ||
<td > | <td > | ||
<B>20</B><br><br><b>Cultivation of</b> | <B>20</B><br><br><b>Cultivation of</b> | ||
| − | OmpA[J23101+B0034+OmpA] A<sup>r</sup> | + | <br>OmpA[J23101+B0034+OmpA] A<sup>r</sup> |
| − | scFv-B[scFv-B+pSB1K3] K<sup>r</sup> | + | <br>scFv-B[scFv-B+pSB1K3] K<sup>r</sup> |
| − | scFv-C[scFv-C+pSB1K3] K<sup>r</sup> | + | <br>scFv-C[scFv-C+pSB1K3] K<sup>r</sup> |
| − | + | <br> | |
<b>Plasmid purification of</b> | <b>Plasmid purification of</b> | ||
| − | OmpA[J23101+B0034+OmpA] A<sup>r</sup> | + | <br>OmpA[J23101+B0034+OmpA] A<sup>r</sup> |
| − | GBP[J23101+B0034+FadL-GBP+pSB1K3] K<sup>r</sup> | + | <br>GBP[J23101+B0034+FadL-GBP+pSB1K3] K<sup>r</sup> |
| − | scFv-B[scFv-B+pSB1K3] K<sup>r</sup> | + | <br>scFv-B[scFv-B+pSB1K3] K<sup>r</sup> |
| − | scFv-C[scFv-C+pSB1K3] K<sup>r</sup> | + | <br>scFv-C[scFv-C+pSB1K3] K<sup>r</sup> |
| − | + | <br> | |
<b>Digestion of </b> | <b>Digestion of </b> | ||
| − | OmpA[J23101+B0034+OmpA] A<sup>r</sup> (EcoRI and Pst1) | + | <br>OmpA[J23101+B0034+OmpA] A<sup>r</sup> (EcoRI and Pst1) |
| − | GBP[J23101+B0034+FadL-GBP+pSB1K3] K<sup>r</sup> (EcoRI and PstI) | + | <br>GBP[J23101+B0034+FadL-GBP+pSB1K3] K<sup>r</sup> (EcoRI and PstI) |
| − | OmpA[J23101+B0034+OmpA] A<sup>r</sup> (NcoI and PstI) | + | <br>OmpA[J23101+B0034+OmpA] A<sup>r</sup> (NcoI and PstI) |
| − | scFv-B[scFv-B] A<sup>r</sup> (EcoRI and PstI) | + | <br>scFv-B[scFv-B] A<sup>r</sup> (EcoRI and PstI) |
| − | scFv-C[scFv-C] A<sup>r</sup> (EcoRI and PstI) | + | <br>scFv-C[scFv-C] A<sup>r</sup> (EcoRI and PstI) |
| − | + | <br> | |
<b>Ligation of</b> | <b>Ligation of</b> | ||
| − | GBP[J23101+B0034+FadL-GBP+pSB1K3] K<sup>r</sup> | + | <br>GBP[J23101+B0034+FadL-GBP+pSB1K3] K<sup>r</sup> |
<b>Transformation off</b> | <b>Transformation off</b> | ||
| − | [J23110+B0034+pSB1K3] K<sup>r</sup> | + | <br>[J23110+B0034+pSB1K3] K<sup>r</sup> |
| − | [pSB3C5] 抗性 | + | <br>[pSB3C5] 抗性 |
| − | + | <br> | |
<b>Electrophoresis of</b> | <b>Electrophoresis of</b> | ||
| − | OmpA[J23101+B0034+OmpA] A<sup>r</sup> (EcoRI and PstI) | + | <br>OmpA[J23101+B0034+OmpA] A<sup>r</sup> (EcoRI and PstI) |
</td> | </td> | ||
<td> | <td> | ||
<B>21</B><br><br><b>Digestion of </b> | <B>21</B><br><br><b>Digestion of </b> | ||
| − | scFv-B[scFv-B] A<sup>r</sup> (EcoRI and PstI) | + | <br>scFv-B[scFv-B] A<sup>r</sup> (EcoRI and PstI) |
| − | scFv-C[scFv-C] A<sup>r</sup> (EcoRI and PstI) | + | <br>scFv-C[scFv-C] A<sup>r</sup> (EcoRI and PstI) |
| − | + | <br> | |
<b>Electrophoresis of</b> | <b>Electrophoresis of</b> | ||
| − | OmpA[J23101+B0034+OmpA] A<sup>r</sup> (NcoI and PstI) | + | <br>OmpA[J23101+B0034+OmpA] A<sup>r</sup> (NcoI and PstI) |
| − | scFv-B[scFv-B] A<sup>r</sup> (EcoRI and PstI) | + | <br>scFv-B[scFv-B] A<sup>r</sup> (EcoRI and PstI) |
| − | scFv-C[scFv-C] A<sup>r</sup> (EcoRI and PstI) | + | <br>scFv-C[scFv-C] A<sup>r</sup> (EcoRI and PstI) |
| − | + | <br> | |
<b>Ligation of</b> | <b>Ligation of</b> | ||
| − | scFv-B[scFv-B+pSB1C3] C<sup>r</sup> | + | <br>scFv-B[scFv-B+pSB1C3] C<sup>r</sup> |
| − | scFv-C[scFv-C+pSB1C3] C<sup>r</sup> | + | <br>scFv-C[scFv-C+pSB1C3] C<sup>r</sup> |
| − | scFv-B[scFv-B+pSB1K3] K<sup>r</sup> | + | <br>scFv-B[scFv-B+pSB1K3] K<sup>r</sup> |
| − | scFv-C[scFv-C+pSB1K3] K<sup>r</sup> | + | <br>scFv-C[scFv-C+pSB1K3] K<sup>r</sup> |
</td> | </td> | ||
<td> | <td> | ||
<B>22</B><br><br><b>Cultivation of</b> | <B>22</B><br><br><b>Cultivation of</b> | ||
| − | [pSB3C5] C<sup>r</sup> | + | <br>[pSB3C5] C<sup>r</sup> |
| − | [J23110+B0034+pSB1K3] K<sup>r</sup> | + | <br>[J23110+B0034+pSB1K3] K<sup>r</sup> |
| − | + | <br> | |
<b>Transformation off</b> | <b>Transformation off</b> | ||
| − | GBP[J23110+B0034+GBP] A<sup>r</sup> | + | <br>GBP[J23110+B0034+GBP] A<sup>r</sup> |
| − | + | <br> | |
<b>PCR of</b> | <b>PCR of</b> | ||
| − | scFv-B[scFv-B+pSB1C3] C<sup>r</sup> | + | <br>scFv-B[scFv-B+pSB1C3] C<sup>r</sup> |
| − | scFv-C[scFv-C+pSB1C3] C<sup>r</sup> | + | <br>scFv-C[scFv-C+pSB1C3] C<sup>r</sup> |
| − | scFv-B[scFv-B+pSB1K3] K<sup>r</sup> | + | <br>scFv-B[scFv-B+pSB1K3] K<sup>r</sup> |
| − | scFv-C[scFv-C+pSB1K3] K<sup>r</sup> | + | <br>scFv-C[scFv-C+pSB1K3] K<sup>r</sup> |
| − | + | <br> | |
<b>Electrophoresis of</b> | <b>Electrophoresis of</b> | ||
| − | scFv-B[scFv-B+pSB1C3] C<sup>r</sup> | + | <br>scFv-B[scFv-B+pSB1C3] C<sup>r</sup> |
| − | scFv-C[scFv-C+pSB1C3] C<sup>r</sup> | + | <br>scFv-C[scFv-C+pSB1C3] C<sup>r</sup> |
| − | scFv-B[scFv-B+pSB1K3] K<sup>r</sup> | + | <br>scFv-B[scFv-B+pSB1K3] K<sup>r</sup> |
| − | scFv-C[scFv-C+pSB1K3] K<sup>r</sup> | + | <br>scFv-C[scFv-C+pSB1K3] K<sup>r</sup> |
| − | + | <br> | |
<b>Ligation of</b> | <b>Ligation of</b> | ||
| − | scFv-B[scFv-B+pSB1A3] A<sup>r</sup> | + | <br>scFv-B[scFv-B+pSB1A3] A<sup>r</sup> |
| − | scFv-C[scFv-C+pSB1A3] A<sup>r</sup> | + | <br>scFv-C[scFv-C+pSB1A3] A<sup>r</sup> |
| − | + | <br> | |
<b>Transform of</b> | <b>Transform of</b> | ||
| − | scFv-B[scFv-B+pSB1A3] A<sup>r</sup> | + | <br>scFv-B[scFv-B+pSB1A3] A<sup>r</sup> |
| − | scFv-C[scFv-C+pSB1A3] A<sup>r</sup> | + | <br>scFv-C[scFv-C+pSB1A3] A<sup>r</sup> |
</td> | </td> | ||
<td > | <td > | ||
<B>23</B><br><br><b>Plasmid purification of</b> | <B>23</B><br><br><b>Plasmid purification of</b> | ||
| − | [pSB3C5] C<sup>r</sup> | + | <br>[pSB3C5] C<sup>r</sup> |
| − | [J23110+B0034+pSB1K3] K<sup>r</sup> | + | <br>[J23110+B0034+pSB1K3] K<sup>r</sup> |
| − | + | <br> | |
<b>Digestion of </b> | <b>Digestion of </b> | ||
| − | [pSB3C5] C<sup>r</sup> (EcoRI and PstI) | + | <br>[pSB3C5] C<sup>r</sup> (EcoRI and PstI) |
| − | [J23110+B0034+pSB1K3] K<sup>r</sup> (EcoRI and PstI) | + | <br>[J23110+B0034+pSB1K3] K<sup>r</sup> (EcoRI and PstI) |
| − | + | <br> | |
<b>PCR of</b> | <b>PCR of</b> | ||
| − | GBP[J23101+B0034+FadL-GBP+pSB1K3] K<sup>r</sup> | + | <br>GBP[J23101+B0034+FadL-GBP+pSB1K3] K<sup>r</sup> |
| − | + | <br> | |
<b>Transformation off</b> | <b>Transformation off</b> | ||
| − | scFv-B[scFv-B+pSB1A3] A<sup>r</sup> | + | <br>scFv-B[scFv-B+pSB1A3] A<sup>r</sup> |
| − | scFv-C[scFv-C+pSB1A3] A<sup>r</sup> | + | <br>scFv-C[scFv-C+pSB1A3] A<sup>r</sup> |
</td> | </td> | ||
| Line 1,168: | Line 1,167: | ||
<td > | <td > | ||
<B>24</B><br><br><b>Electrophoresis of</b> digested products: | <B>24</B><br><br><b>Electrophoresis of</b> digested products: | ||
| − | [pSB3C5] C<sup>r</sup> (EcoRI and PstI) | + | <br>[pSB3C5] C<sup>r</sup> (EcoRI and PstI) |
| − | [J23110+B0034+pSB1K3] K<sup>r</sup> (EcoRI and PstI) | + | <br>[J23110+B0034+pSB1K3] K<sup>r</sup> (EcoRI and PstI) |
| − | + | <br> | |
<b>Cultivation of</b> | <b>Cultivation of</b> | ||
| − | PCR products: GBP[J23101+B0034+FadL-GBP+pSB1K3] K<sup>r</sup> | + | <br>PCR products: GBP[J23101+B0034+FadL-GBP+pSB1K3] K<sup>r</sup> |
| − | scFv-B[scFv-B+pSB1C3] C<sup>r</sup> | + | <br>scFv-B[scFv-B+pSB1C3] C<sup>r</sup> |
| − | scFv-C[scFv-C+pSB1A3] A<sup>r</sup> | + | <br>scFv-C[scFv-C+pSB1A3] A<sup>r</sup> |
| − | + | <br> | |
<b>Plasmid purification of</b> | <b>Plasmid purification of</b> | ||
| − | scFv-B[scFv-B+pSB1A3] A<sup>r</sup> | + | <br>scFv-B[scFv-B+pSB1A3] A<sup>r</sup> |
| − | scFv-C[scFv-C+pSB1A3] A<sup>r</sup> | + | <br>scFv-C[scFv-C+pSB1A3] A<sup>r</sup> |
| − | GFP[B0030+E0040+J61048] | + | <br>GFP[B0030+E0040+J61048] |
| − | + | <br> | |
<b>Digestion of </b> | <b>Digestion of </b> | ||
| − | [J23101+B0034] K<sup>r</sup> (EcoRI and SpeI) | + | <br>[J23101+B0034] K<sup>r</sup> (EcoRI and SpeI) |
| − | + | <br> | |
<b>Ligation of</b> | <b>Ligation of</b> | ||
| − | OmpA[J23101+B0034+OmpA+pSB3C5] C<sup>r</sup> | + | <br>OmpA[J23101+B0034+OmpA+pSB3C5] C<sup>r</sup> |
| − | + | <br> | |
<b>PCR of</b> | <b>PCR of</b> | ||
| − | GBP[J23110+B0034+GBP] A<sup>r</sup> by two pA<sup>r</sup>ts | + | <br>GBP[J23110+B0034+GBP] A<sup>r</sup> by two pA<sup>r</sup>ts |
| − | + | <br> | |
<b>Electrophoresis of</b> | <b>Electrophoresis of</b> | ||
| − | GBP[J23110+B0034+GBP] A<sup>r</sup> | + | <br>GBP[J23110+B0034+GBP] A<sup>r</sup> |
</td> | </td> | ||
<td> | <td> | ||
<B>25</B><br><br><b>Transformation off</b> | <B>25</B><br><br><b>Transformation off</b> | ||
| − | OmpA[J23101+B0034+OmpA+pSB3C5] C<sup>r</sup> | + | <br>OmpA[J23101+B0034+OmpA+pSB3C5] C<sup>r</sup> |
| − | + | <br> | |
<b>Cultivation of</b> | <b>Cultivation of</b> | ||
| − | GBP[J23110+B0034+FadL-GBP] A<sup>r</sup> | + | <br>GBP[J23110+B0034+FadL-GBP] A<sup>r</sup> |
| − | + | <br> | |
<b>Digestion of </b> | <b>Digestion of </b> | ||
| − | GBP[B003+FadL-GBP] C<sup>r</sup> (XbaI and PstI) | + | <br>GBP[B003+FadL-GBP] C<sup>r</sup> (XbaI and PstI) |
| − | scFv-B[scFv-B+pSB1C3] C<sup>r</sup> (EcoRI and PstI) | + | <br>scFv-B[scFv-B+pSB1C3] C<sup>r</sup> (EcoRI and PstI) |
| − | scFv-C[scFv-C+pSB1A3] A<sup>r</sup> (EcoRI and PstI) | + | <br>scFv-C[scFv-C+pSB1A3] A<sup>r</sup> (EcoRI and PstI) |
| − | GFP[B0030+E0040+J61048] (XbaI and PstI) | + | <br>GFP[B0030+E0040+J61048] (XbaI and PstI) |
| − | + | <br> | |
<b>Electrophoresis of</b> | <b>Electrophoresis of</b> | ||
| − | GFP[B0030+E0040+J61048] | + | <br>GFP[B0030+E0040+J61048] |
| − | scFv-B[scFv-B+pSB1C3] C<sup>r</sup> (EcoRI and PstI) | + | <br>scFv-B[scFv-B+pSB1C3] C<sup>r</sup> (EcoRI and PstI) |
| − | scFv-C[scFv-C+pSB1A3] A<sup>r</sup> (EcoRI and PstI) | + | <br>scFv-C[scFv-C+pSB1A3] A<sup>r</sup> (EcoRI and PstI) |
| − | GFP[B0030+E0040+J61048] (XbaI and PstI) | + | <br>GFP[B0030+E0040+J61048] (XbaI and PstI) |
| − | + | <br> | |
<b>Cultivation of</b> | <b>Cultivation of</b> | ||
| − | scFv-B[scFv-B+pSB1C3] C<sup>r</sup> (EcoRI and PstI) | + | <br>scFv-B[scFv-B+pSB1C3] C<sup>r</sup> (EcoRI and PstI) |
| − | scFv-C[scFv-C+pSB1A3] A<sup>r</sup> (EcoRI and PstI) | + | <br>scFv-C[scFv-C+pSB1A3] A<sup>r</sup> (EcoRI and PstI) |
| − | GFP[B0030+E0040+J61048] (XbaI and PstI) | + | <br>GFP[B0030+E0040+J61048] (XbaI and PstI) |
</td> | </td> | ||
<td> | <td> | ||
<B>26</B><br><br><b>Ligation of</b> | <B>26</B><br><br><b>Ligation of</b> | ||
| − | OmpA[J23110+B0034+OmpA+pSB1A3] A<sup>r</sup> | + | <br>OmpA[J23110+B0034+OmpA+pSB1A3] A<sup>r</sup> |
| − | GBP[J23110+B0034+GBP+pSB1A3] A<sup>r</sup> | + | <br>GBP[J23110+B0034+GBP+pSB1A3] A<sup>r</sup> |
| − | + | <br> | |
<b>Gel extraction of</b> | <b>Gel extraction of</b> | ||
| − | [J23110] A<sup>r</sup> (SpeI and PstI) | + | <br>[J23110] A<sup>r</sup> (SpeI and PstI) |
| − | + | <br> | |
<b>Plasmid purification of</b> | <b>Plasmid purification of</b> | ||
| − | GBP[J23110+B0034+GBP] A<sup>r</sup> by two pA<sup>r</sup>ts | + | <br>GBP[J23110+B0034+GBP] A<sup>r</sup> by two pA<sup>r</sup>ts |
| − | + | <br> | |
<b>Digestion of </b> | <b>Digestion of </b> | ||
| − | GBP[J23110+B0034+GBP] A<sup>r</sup> by two pA<sup>r</sup>ts (EcoRI and SpeI) | + | <br>GBP[J23110+B0034+GBP] A<sup>r</sup> by two pA<sup>r</sup>ts (EcoRI and SpeI) |
</td> | </td> | ||
<td > | <td > | ||
<B>27</B><br><br><b>PCR of</b> | <B>27</B><br><br><b>PCR of</b> | ||
| − | [J23101+B0034+OmpA+pSB3C5] | + | <br>[J23101+B0034+OmpA+pSB3C5] |
| + | <br> | ||
<b>Electrophoresis of</b> | <b>Electrophoresis of</b> | ||
| − | GBP[J23110+B0034+GBP] A<sup>r</sup> by two pA<sup>r</sup>ts (EcoRI and SpeI) | + | <br>GBP[J23110+B0034+GBP] A<sup>r</sup> by two pA<sup>r</sup>ts (EcoRI and SpeI) |
| − | GBP[B0034+FadL-GBP] C<sup>r</sup> (XbaI and PstI) | + | <br>GBP[B0034+FadL-GBP] C<sup>r</sup> (XbaI and PstI) |
| − | [J23110] A<sup>r</sup> (SpeI and PstI) | + | <br>[J23110] A<sup>r</sup> (SpeI and PstI) |
</td> | </td> | ||
<td> | <td> | ||
<B>28</B><br><br><b>Ligation of</b> | <B>28</B><br><br><b>Ligation of</b> | ||
| − | scFv-B[scFv-B+pSB1A3] A<sup>r</sup> | + | <br>scFv-B[scFv-B+pSB1A3] A<sup>r</sup> |
| − | scFv-C[scFv-C+pSB1A3] A<sup>r</sup> | + | <br>scFv-C[scFv-C+pSB1A3] A<sup>r</sup> |
| − | GBP[J23110+B0034+FadL-GBP+B0030+E0040+J61048] 抗性?? | + | <br>GBP[J23110+B0034+FadL-GBP+B0030+E0040+J61048] 抗性?? |
| − | scFv-B[J23101+B0034+OmpA+scFv-B+pSB1A3] A<sup>r</sup> | + | <br>scFv-B[J23101+B0034+OmpA+scFv-B+pSB1A3] A<sup>r</sup> |
| − | scFv-C[J23101+B0034+OmpA+scFv-C+pSB1A3] A<sup>r</sup> | + | <br>scFv-C[J23101+B0034+OmpA+scFv-C+pSB1A3] A<sup>r</sup> |
| − | + | <br> | |
<b>Transform of</b> | <b>Transform of</b> | ||
| − | scFv-B[J23101+B0034+OmpA+scFv-B+pSB1A3] A<sup>r</sup> | + | <br>scFv-B[J23101+B0034+OmpA+scFv-B+pSB1A3] A<sup>r</sup> |
| − | scFv-C[J23101+B0034+OmpA+scFv-C+pSB1A3] A<sup>r</sup> | + | <br>scFv-C[J23101+B0034+OmpA+scFv-C+pSB1A3] A<sup>r</sup> |
| − | GBP[J23110+B0034+FadL-GBP+B0030+E0040+J61048] | + | <br>GBP[J23110+B0034+FadL-GBP+B0030+E0040+J61048] |
| − | scFv-B[scFv-B+pSB1A3] A<sup>r</sup> | + | <br>scFv-B[scFv-B+pSB1A3] A<sup>r</sup> |
| − | scFv-C[scFv-C+pSB1A3] A<sup>r</sup> | + | <br>scFv-C[scFv-C+pSB1A3] A<sup>r</sup> |
| − | + | <br> | |
<b>PCR of</b> | <b>PCR of</b> | ||
| − | [J23101+B003+OmpA+pSB3C5] + primer(EX-OmpA and OmpA-S) | + | <br>[J23101+B003+OmpA+pSB3C5] + primer(EX-OmpA and OmpA-S) |
| − | OmpA[J23101+B0034+OmpA+pSB3C5] C<sup>r</sup> | + | <br>OmpA[J23101+B0034+OmpA+pSB3C5] C<sup>r</sup> |
| − | + | <br> | |
| − | Plasmid purification in gel slices of PCR products: | + | <br>Plasmid purification in gel slices of PCR products: |
| − | OmpA[EX-OmpA-S] | + | <br>OmpA[EX-OmpA-S] |
| − | + | <br> | |
<b>Gel extraction of</b> | <b>Gel extraction of</b> | ||
| − | OmpA[OmpA] (EcoRI and SpeI) | + | <br>OmpA[OmpA] (EcoRI and SpeI) |
| − | + | <br> | |
<b>Transformation off</b> | <b>Transformation off</b> | ||
| − | OmpA[J23101+B0034+OmpA+pSB3C5] | + | <br>OmpA[J23101+B0034+OmpA+pSB3C5] |
</td> | </td> | ||
<td> | <td> | ||
<B>29</B><br><br><b>Plasmid purification of</b> | <B>29</B><br><br><b>Plasmid purification of</b> | ||
| − | [J23110] A<sup>r</sup> | + | <br>[J23110] A<sup>r</sup> |
| − | [pSB1C3] | + | <br>[pSB1C3] |
| − | + | <br> | |
<b>Digestion of </b> | <b>Digestion of </b> | ||
| − | [J23110] A<sup>r</sup> (EcoRI and SpeI) | + | <br>[J23110] A<sup>r</sup> (EcoRI and SpeI) |
| − | [pSB1C3] (EcoRI and SpeI) | + | <br>[pSB1C3] (EcoRI and SpeI) |
| − | + | <br> | |
<b>Ligation of</b> | <b>Ligation of</b> | ||
| − | scFv-B[J23101+B0034+OmpA+scFv-B+pSB1A3] A<sup>r</sup> | + | <br>scFv-B[J23101+B0034+OmpA+scFv-B+pSB1A3] A<sup>r</sup> |
| − | scFv-C[J23101+B0034+OmpA+scFv-C+pSB1A3] A<sup>r</sup> | + | <br>scFv-C[J23101+B0034+OmpA+scFv-C+pSB1A3] A<sup>r</sup> |
| − | [J23110+B0034+pSB1K3] K<sup>r</sup> | + | <br>[J23110+B0034+pSB1K3] K<sup>r</sup> |
| − | + | <br> | |
<b>Electrophoresis of</b> | <b>Electrophoresis of</b> | ||
| − | [J23110] A<sup>r</sup> (EcoRI and SpeI) | + | <br>[J23110] A<sup>r</sup> (EcoRI and SpeI) |
| − | [pSB1C3] (EcoRI and SpeI) | + | <br>[pSB1C3] (EcoRI and SpeI) |
| − | + | <br> | |
<b>Transform of</b> | <b>Transform of</b> | ||
| − | scFv-B[J23101+B0034+OmpA+scFv-B+pSB1A3] A<sup>r</sup> | + | <br>scFv-B[J23101+B0034+OmpA+scFv-B+pSB1A3] A<sup>r</sup> |
| − | scFv-C[J23101+B0034+OmpA+scFv-C+pSB1A3] A<sup>r</sup> | + | <br>scFv-C[J23101+B0034+OmpA+scFv-C+pSB1A3] A<sup>r</sup> |
| − | [J23110+B0034+pSB1K3] K<sup>r</sup> | + | <br>[J23110+B0034+pSB1K3] K<sup>r</sup> |
| − | GFP[J23110+B0034+E0040+J61048+pSB1C3] C<sup>r</sup> | + | <br>GFP[J23110+B0034+E0040+J61048+pSB1C3] C<sup>r</sup> |
</td> | </td> | ||
<td > | <td > | ||
<B>30</B><br><br><b>PCR of</b> | <B>30</B><br><br><b>PCR of</b> | ||
| − | scFv-B[J23101+B0034+OmpA+scFv-B+pSB1A3] A<sup>r</sup> | + | <br>scFv-B[J23101+B0034+OmpA+scFv-B+pSB1A3] A<sup>r</sup> |
| − | scFv-C[J23101+B0034+OmpA+scFv-C+pSB1A3] A<sup>r</sup> | + | <br>scFv-C[J23101+B0034+OmpA+scFv-C+pSB1A3] A<sup>r</sup> |
| − | [J23110+B0034+pSB1K3] K<sup>r</sup> | + | <br>[J23110+B0034+pSB1K3] K<sup>r</sup> |
| − | GFP[J23110+B0034+E0040+J61048+pSB1C3] C<sup>r</sup> | + | <br>GFP[J23110+B0034+E0040+J61048+pSB1C3] C<sup>r</sup> |
</td> | </td> | ||
| Line 1,309: | Line 1,309: | ||
<td > | <td > | ||
<B>31</B><br><br><b>Cultivation of</b> | <B>31</B><br><br><b>Cultivation of</b> | ||
| − | [J23110+B0034] K<sup>r</sup> | + | <br>[J23110+B0034] K<sup>r</sup> |
| − | GFP[J23110+B0034+E0040+J61048+pSB1C3] C<sup>r</sup> | + | <br>GFP[J23110+B0034+E0040+J61048+pSB1C3] C<sup>r</sup> |
| − | scFv-C[J23101+B0034+OmpA+scFv-C+pSB1A3] A<sup>r</sup> | + | <br>scFv-C[J23101+B0034+OmpA+scFv-C+pSB1A3] A<sup>r</sup> |
| − | scFv-B[J23101+B0034+OmpA+scFv-B+pSB1A3] A<sup>r</sup> | + | <br>scFv-B[J23101+B0034+OmpA+scFv-B+pSB1A3] A<sup>r</sup> |
| − | + | <br> | |
<b>Plasmid purification of</b> | <b>Plasmid purification of</b> | ||
| − | [J23110+B0034] K<sup>r</sup> | + | <br>[J23110+B0034] K<sup>r</sup> |
| − | GFP[J23110+B0034+E0040+J61048+pSB1C3] C<sup>r</sup> | + | <br>GFP[J23110+B0034+E0040+J61048+pSB1C3] C<sup>r</sup> |
| − | scFv-C[J23101+B0034+OmpA+scFv-C+pSB1A3] A<sup>r</sup> | + | <br>scFv-C[J23101+B0034+OmpA+scFv-C+pSB1A3] A<sup>r</sup> |
| − | scFv-B[J23101+B0034+OmpA+scFv-B+pSB1A3] A<sup>r</sup> | + | <br>scFv-B[J23101+B0034+OmpA+scFv-B+pSB1A3] A<sup>r</sup> |
| − | + | <br> | |
<b>Digestion of </b> | <b>Digestion of </b> | ||
| − | [J23110+B0034] K<sup>r</sup> (EcoRI and PstI) | + | <br>[J23110+B0034] K<sup>r</sup> (EcoRI and PstI) |
| − | GFP[J23110+B0034+E0040+J61048+pSB1C3] C<sup>r</sup> (EcoRI and PstI) | + | <br>GFP[J23110+B0034+E0040+J61048+pSB1C3] C<sup>r</sup> (EcoRI and PstI) |
| − | scFv-C[J23101+B0034+OmpA+scFv-C+pSB1A3] A<sup>r</sup> (EcoRI and PstI) | + | <br>scFv-C[J23101+B0034+OmpA+scFv-C+pSB1A3] A<sup>r</sup> (EcoRI and PstI) |
| − | scFv-B[J23101+B0034+OmpA+scFv-B+pSB1A3] A<sup>r</sup> (EcoRI and PstI) | + | <br>scFv-B[J23101+B0034+OmpA+scFv-B+pSB1A3] A<sup>r</sup> (EcoRI and PstI) |
| − | [J23110+B0034] (EcoRI and SpeI) K<sup>r</sup> | + | <br>[J23110+B0034] (EcoRI and SpeI) K<sup>r</sup> |
| − | [J23101+B0034] (EcoRI and SpeI) K<sup>r</sup> | + | <br>[J23101+B0034] (EcoRI and SpeI) K<sup>r</sup> |
| − | OmpA[OmpA] C<sup>r</sup> (EcoRI and PstI) | + | <br>OmpA[OmpA] C<sup>r</sup> (EcoRI and PstI) |
| − | [pSB1K3] (EcoRI and PstI) | + | <br>[pSB1K3] (EcoRI and PstI) |
| − | + | <br> | |
<b>Electrophoresis of</b> | <b>Electrophoresis of</b> | ||
| − | [J23110+B0034] K<sup>r</sup> (EcoRI and PstI) | + | <br>[J23110+B0034] K<sup>r</sup> (EcoRI and PstI) |
| − | GFP[J23110+B0034+E0040+J61048+pSB1C3] C<sup>r</sup> (EcoRI and PstI) | + | <br>GFP[J23110+B0034+E0040+J61048+pSB1C3] C<sup>r</sup> (EcoRI and PstI) |
| − | scFv-C[J23101+B0034+OmpA+scFv-C+pSB1A3] A<sup>r</sup> (EcoRI and PstI) | + | <br>scFv-C[J23101+B0034+OmpA+scFv-C+pSB1A3] A<sup>r</sup> (EcoRI and PstI) |
| − | scFv-B[J23101+B0034+OmpA+scFv-B+pSB1A3] A<sup>r</sup> (EcoRI and PstI) | + | <br>scFv-B[J23101+B0034+OmpA+scFv-B+pSB1A3] A<sup>r</sup> (EcoRI and PstI) |
| − | + | <br> | |
| − | + | ||
<b>Ligation of</b> | <b>Ligation of</b> | ||
| − | OmpA[OmpA+pSB1C3] C<sup>r</sup> | + | <br>OmpA[OmpA+pSB1C3] C<sup>r</sup> |
</td> | </td> | ||
Revision as of 08:48, 14 September 2015
Lab note
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1 PCR transform |
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15 Transformation of GBP[FadL-GBP] Cultivation of OmpA[OmpA+pSB1K3] Kr |
16 Cultivation of GBP[FadL-GBP] Plasmid purification of OmpA[OmpA+pSB1K3] Kr Digestion of OmpA[OmpA+pSB1K3] Kr (EcoRI and PstI) |
17 Plasmid purification of GBP[FadL-GBP] Digestion of GBP[FadL-GBP] (EcoRI and PstI) Electrophoresis of digested product: GBP[FadL-GBP] (EcoRI and PstI) |
18 Transformation of ligated product: GBP[FadL-GBP+pSB1C3] Cr |
|
19 Cultivation of ligated product: GBP[FadL-GBP] Cr PCR product: Terminator[J61048] |
20 |
21 Cultivation of GFP[J23101+B0034+E0040+J61048+pSB1A3] Ar OmpA[OmpA+pSB1C3] Cr GBP[FadL-GBP+pSB1C3] Cr |
22 Plasmid purification of GFP[J23101+B0034+E0040+J61048+pSB1A3] Ar OmpA[OmpA+pSB1C3] Cr GBP[FadL-GBP+PSB1C3] Cr Digestion of GFP[J23101+B0034+E0040+J61048] Ar (EcoRI and PstI) OmpA[OmpA] Cr (EcoRI and PstI) GBP[FadL-GBP] Cr (EcoRI and PstI) Electrophoresis of digested product: GFP[J23101+B0034+E0040+J61048] Ar (EcoRI and PstI) OmpA[OmpA] Cr (EcoRI and PstI) GBP[FadL-GBP] Cr (EcoRI and PstI) |
23 Digestion of OmpA[OmpA] Cr (EcoRI and SpeI) GBP[FadL-GBP] Cr (EcoRI and SpeI) Electrophoresis of digested product: OmpA[OmpA] Cr (EcoRI and SpeI) GBP[FadL-GBP] Cr (EcoRI and SpeI) |
24 Digestion of OmpA[OmpA] Cr (XbaI and PstI) GBP[FadL-GBP] Cr (XbaI and PstI) |
25 Electrophoresis of digested product: OmpA[OmpA] Cr (XbaI and PstI) GBP[FadL-GBP] Cr (XbaI and PstI) Digestion of RBS[B0034] Ar (XbaI and PstI) backbone[pSB1A3] Ar (EcoRI and PstI) Terminator[J61048] Ar (XbaI and PstI) Electrophoresis of digested product: RBS[B0034] Ar (XbaI and PstI) backbone[pSB1A3] Ar (EcoRI and PstI) Terminator[J61048] Ar (XbaI and PstI) Ligation of OmpA[J23101+B0034+OmpA+pSB1A3] Ar GBP[GBP+B0034+pSB1K3] Kr OmpA[OmpA+J61048+pSB1K3] Kr |
|
26 Ligation of RFP[E1010+B0034+pSB1K3] Kr Transformation of ligated product: OmpA[J23101+B0034+OmpA+pSB1A3] Ar GBP[FadL-GBP+B0034+pSB1K3] Kr OmpA[OmpA+J61048+pSB1K3] Kr Electrophoresis of digested product: RFP[E1010+J61048+pSB1K3] Kr [J23101+B0034] |
27 Digestion of Terminator[J61048] Ar (XbaI and PstI) PCR of ligated product: OmpA[J23101+B0034+OmpA+pSB1A3] Ar OmpA[OmpA+J61048+pSB1K3] Kr |
28 Electrophoresis of RFP[E1010+J61048+pSB1K3] Kr OmpA[J23101+B0034+OmpA+pSB1A3] Ar Transformation of ligated product: GBP[B0034+FadL-GBP+pSB1A3] Ar OmpA[OmpA+J61048+pSB1K3] Kr PCR of ligated product OmpA[J23101+B0034+OmpA] Ar |
29 Cultivation of RFP[E1010+J61048+pSB1K3] Kr PCR of OmpA[J23101+B0034+OmpA] Ar Electrophoresis of OmpA[J23101+B0034+OmpA] Ar Ligation of GBP[FadL-GBP+J61048+pSB1K3] Kr |
30 Plasmid purification of RFP[E1010+J61048+pSB1K3] Kr Ligation of OmpA[J23101+B0034+OmpA] Ar PCR of OmpA[J23101+B0034+OmpA] Ar Transformation of mini: Terminator[J61048] ligated product: OmpA[J23101+B0034+OmpA] Ar |
| Sun. | Mon. | Tue. | Wed. | Thu. | Fri. | Set. |
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1 PCR of Transformation product: mini: Terminator[J61048] ligated product: OmpA[J23101+B0034+OmpA] Ar GBP[B0034+FadL-GBP+pSB1A3] Ar Electrophoresis of mini: Terminator[J61048] ligated product: OmpA[J23101+B0034+OmpA] Ar |
2 |
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3 Ligation of GBP[J23101+B0034+FadL-GBP+pSB1A3] Ar |
4 Ligation of GBP[J23101+B0034+FadL-GBP+pSB1A3] Ar 5/4 Transformation off ligated products: GBP[J23101+B0034+FadL-GBP+pSB1A3] Ar Digestion of RFP[E1010+J61048] (Kr) (XbaI and PstI) PCR of ligated products: GBP[J23101+B0034+FadL-GBP+pSB1A3] Ar Electrophoresis of PCR products: GBP[J23101+B0034+FadL-GBP+pSB1A3] Ar |
5 Electrophoresis of digested products: RFP[E1010+J61048] (Kr) (XbaI and PstI) OmpA[J23101+B0034+OmpA] Ar (EcoRI and PstI) GBP[FadL-GBP+J61048] Kr (EcoRI and PstI) PCR of OmpA[J23101+B0034+OmpA+pSB1A3] Ar Cultivation of PCR product: RFP[E1010+J61048+pSB1K3] Kr GBP[J23101+B0034+FadL-GBP+pSB1A3] Ar |
6 Plasmid purification of RFP[E1010+J61048+pSB1K3] Kr GBP[J23101+B0034+FadL-GBP+pSB1A3] Ar Digestion of GBP[J23101+B0034+FadL-GBP+pSB1A3] Ar (EcoRI and PstI) Cultivation of PCR product: OmpA[J23101+B0034+OmpA+pSB1A3] Ar PCR of backbone: [pSB1C3] Cr [pSB1A3] Ar [pSB1K3] Kr Electrophoresis of PCR products: OmpA[J23101+B0034+OmpA+pSB1A3] Ar digested products: GBP[J23101+B0034+FadL-GBP+pSB1A3] Ar (EcoRI and PstI) |
7 Digestion of RFP[E1010+J61048+pSB1K3] Kr (EcoRI and PstI) RFP[E1010+J61048+pSB1K3] Kr (XbaI and PstI) [pSB1C3] Cr (EcoRI and PstI) [pSB1A3] Ar (EcoRI and PstI) [pSB1K3] Kr (EcoRI and PstI) OmpA[J23101+B0034+OmpA+pSB1A3] Ar (EcoRI and PstI) Plasmid purification of OmpA[J23101+B0034+OmpA+pSB1A3] Ar [pSB1K3] Kr Electrophoresis of digested products: RFP[E1010+J61048+pSB1K3] Kr (EcoRI and PstI) RFP[E1010+J61048+pSB1K3] Kr (XbaI and PstI) [pSB1C3] Cr (EcoRI and PstI) [pSB1A3] Ar (EcoRI and PstI) [pSB1K3] Kr (EcoRI and PstI) Ligation of RFP[B0034+E1010+J61048+pSB1C3] Cr |
8 Electrophoresis of digested products: OmpA[J23101+B0034+OmpA] Ar (EcoRI and PstI) Transformation off ligated products: RFP[B0034+E1010+J61048+pSB1C3] Cr PCR of GBP[J23101+B0034+FadL-GBP+pSB1A3] Ar Electrophoresis of PCR products: GBP[J23101+B0034+FadL-GBP+pSB1A3] Ar Transformation off GBP[J23101+B0034+FadL-GBP+pSB1A3] Ar [B0030] Ar [J23110] Ar |
9 PCR of GBP[J23101+B0034+FadL-GBP+pSB1A3] Ar [B0030] Ar [J23110] Ar Electrophoresis of PCR products: GBP[J23101+B0034+FadL-GBP+pSB1A3] Ar [B0030] Ar [J23110] Ar Cultivation of [B0030] Ar [J23110] Ar Ligation of GBP[J23101+B0034+FadL-GBP+pSB1A3] Ar |
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10 Electrophoresis of [B0030] Ar Cultivation of [B0030] Ar Plasmid purification of [J23110] Ar [B0030] Ar GBP[J23101+B0034+FadL-GBP+pSB1A3] Ar Digestion of [B0030] Ar (EcoRI and SpeI) [J23110] Ar (EcoRI and SpeI) GBP[J23101+B0034+FadL-GBP+pSB1A3] Ar (EcoRI and PstI) GBP[B0034+FadL-GBP+pSB1A3] Ar (XbaI and PstI) Electrophoresis of digested products: [B0030] Ar (EcoRI and SpeI) GBP[J23101+B0034+FadL-GBP+pSB1A3] Ar (EcoRI and PstI) GBP[B0034+FadL-GBP+pSB1A3] Ar (XbaI and PstI) |
11 Electrophoresis of digested products: [J23110] Ar (EcoRI and SpeI) Ligation of RFP[B0030+E1010+J61048+pSB1C3] Cr GFP[B0030+E0040+J61048+pSB1K3] Kr Transformation off RFP[B0030+E1010+J61048+pSB1C3] Cr GFP[B0030+E0040+J61048+pSB1K3] Kr PCR of RFP[B0030+E1010+J61048+pSB1C3] Cr GFP[B0030+E0040+J61048+pSB1K3] Kr |
12 PCR of ligated products: RFP[B0030+E1010+J61048+pSB1C3] Cr Cultivation of PCR products: RFP[B0030+E1010+J61048+pSB1C3] Cr Plasmid purification of RFP[B0030+E1010+J61048+pSB1C3] Cr Digestion of RFP[B0030+E1010+J61048] Cr (EcoRI and SpeI) RFP[B0030+E1010+J61048] Cr (EcoRI and PstI) Electrophoresis of digested products: RFP[B0030+E1010+J61048] Cr (EcoRI and SpeI) RFP[B0030+E1010+J61048] Cr (EcoRI and PstI) PCR products: RFP[B0030+E1010+J61048+pSB1C3] Cr |
13 Ligation of [J23110+B0034+pSB1K3] Kr |
14 Cultivation of OmpA[J23101+B0034+OmpA+pSB1A3] Ar Transformation off [J23110+B0034+pSB1K3] Kr Electrophoresis of digested products: GBP[J23101+B0034+FadL-GBP+pSB1A3] Ar (EcoRI and PstI) GBP[B0034+FadL-GBP+pSB1A3] Ar (XbaI and PstI) [pSB1K3] Kr (EcoRI and PstI) |
15 Digestion of GBP[B0034+FadL-GBP+pSB1A3] Ar (EcoRI and PstI) Ligation of GBP[J23101+B0034+FadL-GBP+pSB1K3] Kr |
16 Transformation off GBP[J23101+B0034+FadL-GBP+pSB1K3] Kr Electrophoresis of GBP[B0034+FadL-GBP+pSB1A3] Ar (EcoRI and PstI) GBP[B0034+FadL-GBP+pSB1A3] Ar (XbaI and PstI) Ligation of GBP[B0034+FadL-GBP+pSB1C3] Cr Transformation off [J23110+B0034+pSB1K3] Kr |
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17 Ligation of [J23110+B0034+pSB1K3] Kr GBP[J23110+B0034+FadL-GBP+pSB1K3] Kr PCR of GBP[B0034+FadL-GBP+pSB1C3] Cr GBP[J23110+B0034+FadL-GBP+pSB1K3] Kr Electrophoresis of PCR products: GBP[B0034+FadL-GBP+pSB1C3] Cr GBP[J23110+B0034+FadL-GBP+pSB1K3] Kr |
18 Cultivation of GBP[B0034+FadL-GBP+pSB1C3] Cr Plasmid purification of GBP[B0034+FadL-GBP+pSB1C3] Cr Digestion of GBP[B0034+FadL-GBP] Cr (XbaI and PstI) RFP[B0030+E1010+J61048] Cr (XbaI and PstI) GFP[B0030+E0040+J61048] Transformation off [J23110+B0034+pSB1K3] Kr GBP[J23101+B0034+FadL-GBP+pSB1K3] Kr GFP[B0030+E0040+J61048] |
19 Ligation of GBP[J23101+B0034+FadL-GBP+pSB1K3] Kr PCR of GBP[J23101+B0034+FadL-GBP+pSB1K3] Kr Electrophoresis of digested products: RFP[B0030+E1010+J61048] Cr (XbaI and PstI) GBP[B0034+FadL-GBP] Cr (XbaI and PstI) PCR products: GBP[J23101+B0034+FadL-GBP+pSB1K3] Kr Transformation off scFv-B[scFv-B+pSB1K3] Kr scFv-C[scFv-C+pSB1K3] Kr GFP[B0030+E0040+J61048] Cultivation of GBP[J23101+B0034+FadL-GBP+pSB1K3] Kr |
20 Cultivation of OmpA[J23101+B0034+OmpA] Ar scFv-B[scFv-B+pSB1K3] Kr scFv-C[scFv-C+pSB1K3] Kr Plasmid purification of OmpA[J23101+B0034+OmpA] Ar GBP[J23101+B0034+FadL-GBP+pSB1K3] Kr scFv-B[scFv-B+pSB1K3] Kr scFv-C[scFv-C+pSB1K3] Kr Digestion of OmpA[J23101+B0034+OmpA] Ar (EcoRI and Pst1) GBP[J23101+B0034+FadL-GBP+pSB1K3] Kr (EcoRI and PstI) OmpA[J23101+B0034+OmpA] Ar (NcoI and PstI) scFv-B[scFv-B] Ar (EcoRI and PstI) scFv-C[scFv-C] Ar (EcoRI and PstI) Ligation of GBP[J23101+B0034+FadL-GBP+pSB1K3] Kr Transformation off [J23110+B0034+pSB1K3] Kr [pSB3C5] 抗性 Electrophoresis of OmpA[J23101+B0034+OmpA] Ar (EcoRI and PstI) |
21 Digestion of scFv-B[scFv-B] Ar (EcoRI and PstI) scFv-C[scFv-C] Ar (EcoRI and PstI) Electrophoresis of OmpA[J23101+B0034+OmpA] Ar (NcoI and PstI) scFv-B[scFv-B] Ar (EcoRI and PstI) scFv-C[scFv-C] Ar (EcoRI and PstI) Ligation of scFv-B[scFv-B+pSB1C3] Cr scFv-C[scFv-C+pSB1C3] Cr scFv-B[scFv-B+pSB1K3] Kr scFv-C[scFv-C+pSB1K3] Kr |
22 Cultivation of [pSB3C5] Cr [J23110+B0034+pSB1K3] Kr Transformation off GBP[J23110+B0034+GBP] Ar PCR of scFv-B[scFv-B+pSB1C3] Cr scFv-C[scFv-C+pSB1C3] Cr scFv-B[scFv-B+pSB1K3] Kr scFv-C[scFv-C+pSB1K3] Kr Electrophoresis of scFv-B[scFv-B+pSB1C3] Cr scFv-C[scFv-C+pSB1C3] Cr scFv-B[scFv-B+pSB1K3] Kr scFv-C[scFv-C+pSB1K3] Kr Ligation of scFv-B[scFv-B+pSB1A3] Ar scFv-C[scFv-C+pSB1A3] Ar Transform of scFv-B[scFv-B+pSB1A3] Ar scFv-C[scFv-C+pSB1A3] Ar |
23 Plasmid purification of [pSB3C5] Cr [J23110+B0034+pSB1K3] Kr Digestion of [pSB3C5] Cr (EcoRI and PstI) [J23110+B0034+pSB1K3] Kr (EcoRI and PstI) PCR of GBP[J23101+B0034+FadL-GBP+pSB1K3] Kr Transformation off scFv-B[scFv-B+pSB1A3] Ar scFv-C[scFv-C+pSB1A3] Ar |
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24 Electrophoresis of digested products: [pSB3C5] Cr (EcoRI and PstI) [J23110+B0034+pSB1K3] Kr (EcoRI and PstI) Cultivation of PCR products: GBP[J23101+B0034+FadL-GBP+pSB1K3] Kr scFv-B[scFv-B+pSB1C3] Cr scFv-C[scFv-C+pSB1A3] Ar Plasmid purification of scFv-B[scFv-B+pSB1A3] Ar scFv-C[scFv-C+pSB1A3] Ar GFP[B0030+E0040+J61048] Digestion of [J23101+B0034] Kr (EcoRI and SpeI) Ligation of OmpA[J23101+B0034+OmpA+pSB3C5] Cr PCR of GBP[J23110+B0034+GBP] Ar by two pArts Electrophoresis of GBP[J23110+B0034+GBP] Ar |
25 Transformation off OmpA[J23101+B0034+OmpA+pSB3C5] Cr Cultivation of GBP[J23110+B0034+FadL-GBP] Ar Digestion of GBP[B003+FadL-GBP] Cr (XbaI and PstI) scFv-B[scFv-B+pSB1C3] Cr (EcoRI and PstI) scFv-C[scFv-C+pSB1A3] Ar (EcoRI and PstI) GFP[B0030+E0040+J61048] (XbaI and PstI) Electrophoresis of GFP[B0030+E0040+J61048] scFv-B[scFv-B+pSB1C3] Cr (EcoRI and PstI) scFv-C[scFv-C+pSB1A3] Ar (EcoRI and PstI) GFP[B0030+E0040+J61048] (XbaI and PstI) Cultivation of scFv-B[scFv-B+pSB1C3] Cr (EcoRI and PstI) scFv-C[scFv-C+pSB1A3] Ar (EcoRI and PstI) GFP[B0030+E0040+J61048] (XbaI and PstI) |
26 Ligation of OmpA[J23110+B0034+OmpA+pSB1A3] Ar GBP[J23110+B0034+GBP+pSB1A3] Ar Gel extraction of [J23110] Ar (SpeI and PstI) Plasmid purification of GBP[J23110+B0034+GBP] Ar by two pArts Digestion of GBP[J23110+B0034+GBP] Ar by two pArts (EcoRI and SpeI) |
27 PCR of [J23101+B0034+OmpA+pSB3C5] Electrophoresis of GBP[J23110+B0034+GBP] Ar by two pArts (EcoRI and SpeI) GBP[B0034+FadL-GBP] Cr (XbaI and PstI) [J23110] Ar (SpeI and PstI) |
28 Ligation of scFv-B[scFv-B+pSB1A3] Ar scFv-C[scFv-C+pSB1A3] Ar GBP[J23110+B0034+FadL-GBP+B0030+E0040+J61048] 抗性?? scFv-B[J23101+B0034+OmpA+scFv-B+pSB1A3] Ar scFv-C[J23101+B0034+OmpA+scFv-C+pSB1A3] Ar Transform of scFv-B[J23101+B0034+OmpA+scFv-B+pSB1A3] Ar scFv-C[J23101+B0034+OmpA+scFv-C+pSB1A3] Ar GBP[J23110+B0034+FadL-GBP+B0030+E0040+J61048] scFv-B[scFv-B+pSB1A3] Ar scFv-C[scFv-C+pSB1A3] Ar PCR of [J23101+B003+OmpA+pSB3C5] + primer(EX-OmpA and OmpA-S) OmpA[J23101+B0034+OmpA+pSB3C5] Cr Plasmid purification in gel slices of PCR products: OmpA[EX-OmpA-S] Gel extraction of OmpA[OmpA] (EcoRI and SpeI) Transformation off OmpA[J23101+B0034+OmpA+pSB3C5] |
29 Plasmid purification of [J23110] Ar [pSB1C3] Digestion of [J23110] Ar (EcoRI and SpeI) [pSB1C3] (EcoRI and SpeI) Ligation of scFv-B[J23101+B0034+OmpA+scFv-B+pSB1A3] Ar scFv-C[J23101+B0034+OmpA+scFv-C+pSB1A3] Ar [J23110+B0034+pSB1K3] Kr Electrophoresis of [J23110] Ar (EcoRI and SpeI) [pSB1C3] (EcoRI and SpeI) Transform of scFv-B[J23101+B0034+OmpA+scFv-B+pSB1A3] Ar scFv-C[J23101+B0034+OmpA+scFv-C+pSB1A3] Ar [J23110+B0034+pSB1K3] Kr GFP[J23110+B0034+E0040+J61048+pSB1C3] Cr |
30 PCR of scFv-B[J23101+B0034+OmpA+scFv-B+pSB1A3] Ar scFv-C[J23101+B0034+OmpA+scFv-C+pSB1A3] Ar [J23110+B0034+pSB1K3] Kr GFP[J23110+B0034+E0040+J61048+pSB1C3] Cr |
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31 Cultivation of [J23110+B0034] Kr GFP[J23110+B0034+E0040+J61048+pSB1C3] Cr scFv-C[J23101+B0034+OmpA+scFv-C+pSB1A3] Ar scFv-B[J23101+B0034+OmpA+scFv-B+pSB1A3] Ar Plasmid purification of [J23110+B0034] Kr GFP[J23110+B0034+E0040+J61048+pSB1C3] Cr scFv-C[J23101+B0034+OmpA+scFv-C+pSB1A3] Ar scFv-B[J23101+B0034+OmpA+scFv-B+pSB1A3] Ar Digestion of [J23110+B0034] Kr (EcoRI and PstI) GFP[J23110+B0034+E0040+J61048+pSB1C3] Cr (EcoRI and PstI) scFv-C[J23101+B0034+OmpA+scFv-C+pSB1A3] Ar (EcoRI and PstI) scFv-B[J23101+B0034+OmpA+scFv-B+pSB1A3] Ar (EcoRI and PstI) [J23110+B0034] (EcoRI and SpeI) Kr [J23101+B0034] (EcoRI and SpeI) Kr OmpA[OmpA] Cr (EcoRI and PstI) [pSB1K3] (EcoRI and PstI) Electrophoresis of [J23110+B0034] Kr (EcoRI and PstI) GFP[J23110+B0034+E0040+J61048+pSB1C3] Cr (EcoRI and PstI) scFv-C[J23101+B0034+OmpA+scFv-C+pSB1A3] Ar (EcoRI and PstI) scFv-B[J23101+B0034+OmpA+scFv-B+pSB1A3] Ar (EcoRI and PstI) Ligation of OmpA[OmpA+pSB1C3] Cr |
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1 Electrophoresis of [J23110+B0034] (EcoRI and SpeI) Kr [J23101+B0034] (EcoRI and SpeI) Kr OmpA[OmpA] (NcoI and PstI) GFP[K1489002+E0040] Kr Digestion of [pSB1C3] (EcoRI and PstI) [pSB1A3] (EcoRI and PstI) OmpA[OmpA] (EcoRI and NcoI) [pSB3C5] (EcoRI and PstI) Ligation of GFP[J23101+B0034+K1489002+E0040+pSB3C5] (Cr) GFP[J23110+B0034+K1489002+E0040+pSB3C5] (Cr) Electrophoresis of digested product: [pSB1C3] (EcoRI and PstI) [pSB1A3] (EcoRI and PstI) OmpA[OmpA] (EcoRI and NcoI) Cultivation of BFP[OmpA+K592100] PCR of GBP[J23101+B0034+FadL-GBP+pSB1K3] Kr Electrophoresis of GBP[J23101+B0034+FadL-GBP+pSB1K3] Kr |
2 PCR of OmpA[EX-OmpA-S+pSB1C3] Cr Electrophoresis of OmpA[EX-OmpA-S+pSB1C3] Cr OmpA[OmpA] (EcoRI and NcoI) Ligation of scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E1010+J61048+pSB1K3] Kr scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E1010+J61048+pSB1K3] Kr Digestion of [pSB1A3] (EcoRI and NcoI) Electrophoresis of digested products: [pSB1A3] (EcoRI and NcoI) |
3 Ligation of OmpA[OmpA+psB1C3] Cr [J23110+B0034+pSB1K3] Kr Transform of OmpA[OmpA+psB1C3] Cr [J23110+B0034+pSB1K3] Kr Digestion of BFP[OmpA+ K592100] (NcoI and PstI) OmpA[OmpA] (NcoI and PstI) |
4 Cultivation of [J23110+pSB1A3] Ar Transform of [pSB1C3] Digestion of [pSB1C3] (EcoRI and PstI) Ligation of OmpA[OmpA+pSB1C3] Cr OmpA[J23101+B0034+OmpA+pSB3C5] Cr Transform of [B0034+pSB1A3] Ar |
5 Transform of OmpA[J23101+B0034+OmpA+pSB3C5] Cr Plasmid purification of [J23110+pSb1A3] Ar Digestion of [J23110+pSb1A3] Ar PCR of [J23110+pSb1A3] Ar BFP[J23101+B0034+OmpA+K592100+pSB3C5] Cr Transform of OmpA[OmpA+pSB1C3] Cr Electrophoresis of [pSB1C3] [B0034+pSB1A3] Ar BFP[J23101+B0034+OmpA+K592100+pSB3C5] Cr |
6 Cultivation of [pSB1C3] Digestion of [pSB1C3] (EcoRI and SpeI) |
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7 Gel extraction of OmpA[EX-OmpA-S] 抗性 [pSB1C3] (EcoRI and SpeI) Gel extraction of [pSB1C3] (EcoRI and SpeI) Transform of GFP[J23101+B0034+E0040+J61048+pSB1A3] Ar [pSB1A3] OmpA[J23101+B0034+OmpA] Ar Ligation of OmpA[OmpA+pSB1C3] Cr Cultivation of [pSB1C3] |
8 Transform of OmpA[OmpA+pSB1C3] Cr [pSB3K3] [pSB6A1] Plasmid purification of [pSB1C3] Digestion of OmpA[EX-OmpA-S] (EcoRI and SpeI) Electrophoresis of [pSB1C3] (EcoRI and SpeI) Transform of GBP[J23110+B0034+FadL-GBP+B0030+E0040+J61048+pSB1K3] Kr |
9 Ligation of OmpA[OmpA+pSB1C3] Cr PCR of GBP[J23110+B0034+FadL-GBP+B0030+E0040+J61048+pSB1K3] Kr Cultivation of BFP[J23101+B0034+OmpA+K592100+pSB3C5] Cr Plasmid purification of [pSB3K3] [pSB6A1] Digestion of [pSB3K3] (EcoRI and PstI) [pSB6A1] (EcoRI and PstI) |
10 Transform of OmpA[OmpA+pSB1C3] Cr Plasmid purification of [J23101+B0034+OmpA+K592100+pSB3C5] Cr Digestion of [J23101+B0034+OmpA+K592100] Cr (EcoRI and PstI) Electrophoresis of [pSB3K3] (EcoRI and PstI) [pSB6A1] (EcoRI and PstI) [pSB1A3] (EcoRI and PstI) scFv-B[J23101+B0034+OmpA+scFv-B] (EcoRI and PstI) scFv-C[J23101+B0034+OmpA+scFv-C] (EcoRI and PstI) Transform of [J23110+B0034] Kr scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB1C3] scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB1C3] Ligation of scFv-B[J23101+B0034+OmpA+scFv-B+pSB3C5] Cr scFv-C[J23101+B0034+OmpA+scFv-C+pSB3C5] Cr Electrophoresis of [pSB3K3] BFP[J23101+B0034+OmpA+K592100+pSB3C5] Cr |
11 Transform of scFv-B[J23101+B0034+OmpA+scFv-B+pSB3C5] Cr scFv-C[J23101+B0034+OmpA+scFv-C+pSB3C5] Cr [J23110+B0034+pSB1C3] Cr Ligation of [J23110+B0034+pSB1K3] Kr [J23110+B0034+pSB1C3] Cr PCR of OmpA[OmpA+pSB1C3] Cr scFv-B[J23101+B0034+OmpA+scFv-B+pSB3C5] Cr scFv-C[J23101+B0034+OmpA+scFv-C+pSB3C5] Cr Electrophoresis of OmpA[OmpA+pSB1C3] Cr scFv-B[J23101+B0034+OmpA+scFv-B+pSB3C5] Cr scFv-C[J23101+B0034+OmpA+scFv-C+pSB3C5] Cr Cultivation of OmpA[OmpA+pSB1C3] Cr |
12 PCR of [J23110+B0034+pSB1C3] Cr [B0034+pSB1A3] Ar Plasmid purification of OmpA[OmpA+pSB1C3] Cr Ligation of [J23110+B0034+pSB1C3] Cr Transform of [J23110+B0034+pSB1C3] Cr |
13 Electrophoresis of scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB1C3] Cr (EcoRI and PstI) scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB1C3] Cr (EcoRI and PstI) |
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14 SDS-page of [pSB1C3] scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB1C3] Cr (EcoRI and PstI) scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB1C3] Cr (EcoRI and PstI) [pSB3C5] scFv-B[J23101+B0034+OmpA+scFv-B+pSB3C5] Cr scFv-C[J23101+B0034+OmpA+scFv-C+pSB3C5] Cr scFv-B[OmpA+scFv-B] scFv-C[OmpA+scFv-C] Electrophoresis of [J23110] [B0034+FadL-GBP+B0030+E1010+J61048] 抗性 (XbaI and PstI) Ligation of GBP[J23110+B0034+FadL-GBP+B0030+E0040+J61048+pSB3K3] Cr GBP[J23110+B0034+FadL-GBP+B0030+E0040+J61048+pSB1K3] Cr Digestion of OmpA[OmpA+pSB1C3] Cr (EcoRI and SpeI) OmpA[OmpA+pSB1C3] Cr (EcoRI and PstI) Transform of scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB6A1] Ar scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB6A1] Ar Ligation of GBP[FadL-GBP+B0030+E0040+J61048+pSB6A1] Ar Transform of GBP[FadL-GBP+B0030+E0040+J61048+pSB6A1] Ar |
15 |
16 Transform of [pSB3C5] |
17 Cultivation of [pSB3C5] |
18 Plasmid purification of [pSB3C5] Digestion of [pSB3C5]( EcoRI and PstI) Electrophoresis of [pSB3C5]( EcoRI and PstI) Ligation of BFP[J23101+B0034+OmpA+K592100+pSB3C5] Cr BFP[J23101+B0034+OmpA+K592100+pSB1C3] Cr |
19 Transform of BFP[J23101+B0034+OmpA+K592100+pSB3C5] Cr BFP[J23101+B0034+OmpA+K592100+pSB1C3] Cr |
20 PCR of BFP[J23101+B0034+OmpA+K592100+pSB3C5] Cr BFP[J23101+B0034+OmpA+K592100+pSB1C3] Cr Electrophoresis of BFP[J23101+B0034+OmpA+K592100+pSB3C5] Cr BFP[J23101+B0034+OmpA+K592100+pSB1C3] Cr |
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21 |
22 |
23 PCR of GBP[J23110+B0034+FadL-GBP+B0030+E0040+J61048+pSB1K3] Kr Cultivation of [pSB3C5] scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB6A1] Ar BFP[OmpA+K592100] 抗性 scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB6A1] [psB6A1] GBP[J23110+B0034+FadL-GBP+B0030+E0040+J61048+pSB6A1] Ar [pSB1A3] GBP[J23110+B0034+FadL-GBP+B0030+E0040+J61048+pSB1C3] Cr Ligation of scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB6A1] Ar |
24 Plasmid purification of [pSB3C5] |
25 Cultivation of scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB1C3] Cr Transform of [J23101+B0034+pSB1K3] Kr |
26 |
27 Digestion of OmpA[OmpA] Cr (EcoRI and PstI) Electrophoresis of GBP[FadL-GBP] 抗性 (EcoRI and PstI) scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048] Ar (EcoRI and PstI) OmpA[OmpA] Cr (EcoRI and PstI) |
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28 Cultivation of BFP[J23101+B0034+OmpA+K592100+pSB1A3] Ar GBP[J23110+B0034+FadL-GBP+B0030+E0040+J61048+pSB3C5] Cr GBP[J23110+B0034+FadL-GBP+B0030+E0040+J61048+pSB6A1] Ar scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB1C3] Cr [pSB6A1] [pSB3C5] [pSB1C3] [pSB1A3] scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB1C3] Cr Electrophoresis of [J23101+B0034] Kr GBP[FadL-GBP] 抗性 (EcoRI and PstI) scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048] Ar (EcoRI and PstI) OmpA[OmpA] Cr (EcoRI and PstI) BFP[J23101+B0034+OmpA+K592100] Ar (EcoRI and PstI) scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048] Cr (EcoRI and PstI) scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048] Cr (EcoRI and PstI) [pSB6A1] (EcoRI and PstI) OmpA[OmpA] Cr (XbaI and PstI) [E0040] Ar (XbaI and PstI) Ligation of scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB6A1] Ar scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB6A1] Ar BFP[J23101+B0034+OmpA+K592100+pSB3C5] Cr [B0034+E0040+pSB1K3] Kr GBP[FadL-GBP+pSB6A1] Ar GBP[FadL-GBP+pSB1C3] Cr GBP[FadL-GBP+pSB3C5] Cr Digestion of BFP[J23101+B0034+OmpA+K592100] Cr (XbaI and PstI) SDS-page of [pSB1A3] [pSB1C3] BFP[J23101+B0034+OmpA+K592100+pSB1A3] Ar scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB1C3] Cr scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB1C3] Cr [pSB3C5] [pSB6A1] GBP[J23110+B0034+FadL-GBP+B0030+E0040+J61048+pSB3C5] Cr GBP[J23110+B0034+FadL-GBP+B0030+E0040+J61048+pSB6A1] Ar Transform of scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB6A1] Ar scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB6A1] Ar GBP[FadL-GBP+pSB1C3] Cr GBP[FadL-GBP+pSB3C5] Cr BFP[J23101+B0034+OmpA+K592100+pSB3C5] Cr |
29 PCR of scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB6A1] Ar scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB6A1] Ar GBP[FadL-GBP+pSB1C3] Cr GBP[FadL-GBP+pSB3C5] Cr BFP[J23101+B0034+OmpA+K592100+pSB3C5] Cr Electrophoresis of scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB6A1] Ar scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB6A1] Ar |
30 Electrophoresis of scFv-C[J23101+B0034+scFv-C+B0030+E0040+J61048+pSB6A1] Ar scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB6A1] Ar |
| Sun. | Mon. | Tue. | Wed. | Thu. | Fri. | Set. |
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1 Cultivation of [pSB6A1] scFv-C[J23101+B0034+scFv-C+B0030+E0040+J61048+pSB6A1] Ar scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB6A1] Ar Transform of scFv-C[J23101+B0034+scFv-C+B0030+E0040+J61048+pSB6A1] Ar scFv-B[J23101+B0034+scFv-B+B0030+E0040+J61048+pSB6A1] Ar scFv-B[J23101+B0034+OmpA+scFv-B+pSB3C5] Cr scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB1C3] Cr Digestion of GBP[]FadL-GBP] 抗性 (EcoRI and PstI) |
2 Digestion of [J23110] Ar (SpeI and PstI) Plasmid purification in gel slices of scFv-C[J23101+B0034+scFv-C+B0030+E0040+J61048+pSB6A1] Ar scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB6A1] Ar Digestion of scFv-C[J23101+B0034+scFv-C+B0030+E0040+J61048+pSB6A1] Ar (EcoRI and PstI) scFv-B[J23101+B0034+scFv-B+B0030+E0040+J61048+pSB6A1] Ar (EcoRI and PstI) Electrophoresis of scFv-C[J23101+B0034+scFv-C+B0030+E0040+J61048+pSB6A1] Ar (EcoRI and PstI) scFv-B[J23101+B0034+scFv-B+B0030+E0040+J61048+pSB6A1] Ar (EcoRI and PstI) |
3 Gel extraction of [J23110] Ar (SpeI and PstI) [pSB6A1] (EcoRI and PstI) Transform of scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB1C3] Cr Electrophoresis of GBP[FadL-GBP] (EcoRI and Pstl) Ligation of GBP[FadL-GBP+pSB3C5] Cr GBP[FadL-GBP+pSB1C3] Cr GBP[FadL-GBP+pSB6A1] Ar |
4 Transform of GBP[FadL-GBP+pSB3C5] Cr GBP[FadL-GBP+pSB1C3] Cr GBP[FadL-GBP+pSB6A1] Ar scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB1C3] Cr scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB1C3] Cr Electrophoresis of scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB1C3] Cr Cultivation of scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB1C3] Cr [J23101+B0034+pSB1K3] Kr |
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5 Plasmid purification of [J23101+B0034+pSB1K3] Kr scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB1C3] Cr |
6 Cultivation of [pSB1C3] scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB1C3] Cr [pSB6A1] scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB6A1] Ar scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB6A1] Ar [pSB1A3] GFP[J23101+B0034+E0040+J61048+pSB1A3] Ar |
7 Digestion of [J23101+B0034+pSB1K3] Kr (EcoRI and PstI) SDS-page of [pSB1C3] scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB1C3] Cr [pSB6A1] scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB6A1] Ar scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB6A1] Ar [pSB1A3] GFP[J23101+B0034+E0040+J61048+pSB1A3] Ar Ligation of [J23110+B0034] Ar Electrophoresis of [J23101+B0034+pSB1K3] Kr (EcoRI and PstI) scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB6A1] Ar |
8 Plasmid purification of scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB6A1] Ar Transform of [J23110+B0034] Ar BFP[J23101+B0034+OmpA+K592100] scFv-B[J23101+B0034+OmpA+scFv-B+B0030+K592100] Electrophoresis of scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB6A1] Ar (EcoRI and PstI) |
9 Electrophoresis of scFv-C[J23101+B0034+scFv-C+B0030+E0040+J61048+pSB6A1] Ar scFv-B[J23101+B0034+scFv-B+B0030+E0040+J61048+pSB6A1] Ar scFv-B[J23101+B0034+scFv-B+B0034+E0040+J61048+pSB1C3] Cr [pSB3C5] BFP[J23101+B0034+OmpA+scFv-B+B0030+e0040+J61048+pSB6A1] Ar Cultivation of scFv-B[J23110+B0034+scFv-B+B0030+E0040+J61048+pSB6A1] Ar |
10 Plasmid purification of scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB6A1] Ar |
11 PCR of [J23110+B0034] Ar Electrophoresis of [J23110+B0034] Ar |
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12 Cultivation of scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB6A1] Ar scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB6A1] Ar [J23110+B0034] Ar Plasmid purification of scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB6A1] Ar scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB6A1] Ar Digestion of scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB6A1] Ar (EcoRI and PstI) scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB6A1] Ar (EcoRI and PstI) |
13 Plasmid purification of [J23110+B0034] Ar Transform of [R0010] Electrophoresis of scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB6A1] Ar (EcoRI and PstI) scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB6A1] Ar (EcoRI and PstI) |
14 Digestion of [J23110+B0034] Ar (EcoRI and PstI) GBP[FadL-GBP] Kr (XbaI and PstI) OmpA[OmpA] Cr (EcoRI and SpeI) Electrophoresis of [J23110+B0034] Ar (EcoRI and PstI) GBP[FadL-GBP] Kr (XbaI and PstI) OmpA[OmpA] Cr (EcoRI and SpeI) Transform of [R0010] Cr Cultivation of [pSB1C3] scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB1C3] Cr scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB1C3] Cr |
15 Digestion of [pSB6A1] [pSB3C5] Anti-Her 2[Herceptin+E1010] [J23101+B0034+OmpA+scFv-C] [J23101+B0034+OmpA+scFv-B] Cultivation of [pSB6A1] [R0010] PCR of OmpA[OmpA-linker] (EcoRI and PstI) Transform of [J23110+B0034] Kr BFP[K592100+J61048] Kr |
16 Gel extraction of OmpA[OmpA-linker] (EcoRI and PstI) Ligation of OmpA[J23101+B0034+OmpA+pSB3C5] Cr BFP[J23101+B0034+OmpA+K592100+pSB3C5] Cr Digestion of OmpA[OmpA-linker] (EcoRI and PstI) Transform of OmpA[J23101+B0034+OmpA+pSB3C5] Cr |
17 Electrophoresis of OmpA[OmpA-linker] (EcoRI and PstI) Anti-Her 2[Herceptin+E1010] (EcoRI and PstI) scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB6A1] Ar (EcoRI and PstI) scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB6A1] Ar (EcoRI and PstI) |
18 Ligation of OmpA[OmpA-linker+pSB1C3] Cr Anti-Her 2[Herceptin+E1010+pSB1C3] Cr |
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19 Transform of OmpA[OmpA-linker+pSB1C3] Cr Anti-Her 2[Herceptin+E1010+pSB1C3] Cr |
20 |
21 |
22 |
23 |
24 Transform of OmpA[OmpA-linker+pSB1C3] Cr Anti-Her 2[Herceptin+E1010+pSB1C3] Cr PCR of OmpA[OmpA-linker+pSB1C3] Cr Anti-Her 2[Herceptin+E1010+pSB1C3] Cr Electrophoresis of OmpA[OmpA-linker+pSB1C3] Cr Anti-Her 2[Herceptin+E1010+pSB1C3] Cr |
25 Cultivation of [R0010] Cr BFP[K592100+J61048] Kr PCR of OmpA[OmpA-linker+pSB1C3] Cr Anti-Her 2[Herceptin+E1010+pSB1C3] Cr Electrophoresis of OmpA[OmpA-linker+pSB1C3] Cr Anti-Her 2[Herceptin+E1010+pSB1C3] Cr |
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26 Plasmid purification of [R0010] Cr BFP[K592100+J61048] Kr Digestion of [R0010] Cr (EcoRI and SpeI) BFP[K592100+J61048] Kr (EcoRI and PstI) Electrophoresis of [R0010] Cr BFP[K592100+J61048] Kr Ligation of [R0010+B0034+pSB1K3] Kr GBP[R0010+B0034+FadL-GBP+B0030+E0040+J61048+pSB1K3] Kr BFP[B0030+K592100+J61048+pSB1C3] Cr Transform of scFv-C[J23101+B0034+OmpA+scFv-C] Ar Ligation of OmpA[OmpA-linker+pSB1C3] Cr PCR of GBP[J23110+B0034+FadL-GBP+B0030+E0040+J61048] Cultivation of [pSB1C3] GFP[J23101+B0034+E0040+J61048+pSB1A3] Ar scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB1C3] Cr scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB1C3] Cr BFP[J23101+B0034+OmpA+K592100+pSB1C3] Cr GBP[FadL-GBP+pSB6A1] Ar |
27 PCR of GBP[FadL-GBP] Anti-Her 2[Herceptin+E1010] Gel extraction of GBP[FadL-GBP] Anti-Her 2[Herceptin+E1010] Transform of [R0010+B0034+pSB1K3] GBP[R0010+B0034+FadL-GBP+B0030+J61048+pSB1K3] BFP[B0030+K592100+J61048+pSB1C3] Cr PCR of GBP[J23110+B0034+FadL-GBP+E0040+J61048] Cr Gel extraction of GBP[J23110+B0034+FadL-GBP+E0040+J61048] Cr SDS-page of [pSB1C3] GFP[J23101+B0034+E0040+J61048+pSB1C3] Cr scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB1C3] Cr scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB1C3] Cr BFP[J23101+B0034+OmpA+K592100+pSB1C3] Cr GBP[J23110+B0034+FadL-GBP+E0040+J61048+pSB1C3] Cr GBP[FadL-GBP+pSB6A1] Ar [pSB6A1] |
28 Ligation of [R0010+B0034+pSB1K3] GBP[R0010+B0034+FadL-GBP+B0030+E0040+J661048+pSB1K3] Kr BFP[B0030+K592100+J61048+pSB1C3] Cr GBP[R0010+B0034+FadL-GBP+B0030+E0040+J661048+pSB1C3] Cr GBP[R0010+B0034+FadL-GBP+B0030+E0040+J661048+pSB6A1] Ar PCR of BFP[B0030+K592100+J61048+pSB1C3] Cr [R0010+B0034+pSB1K3] GBP[R0010+B0034+FadL-GBP+B0030+E0040+J661048+pSB1K3] Kr OmpA[J23110+B0034+OmpA+pSB3C5] Cr Digestion of GBP[R0010+B0034+FadL-GBP+B0030+E0040+J661048] (XbaI and PstI) Electrophoresis of GBP[R0010+B0034+FadL-GBP+B0030+E0040+J661048] (XbaI and PstI) SDS-page of [pSB1C3] BFP[J23101+B0034+OmpA+K592100+pSB1C3] Cr GFP[J23101+B0034+E0040+J61048+pSB1C3] Cr scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB1C3] Cr scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB1C3] Cr GBP[J23110+B0034+FadL-GBP+B0030+E0040+J61048+pSB1C3] Cr [pSB6A1] GBP[R0010+B0034+FadL-GBP+B0030+E0040+J661048+pSB6A1] Ar Transform of OmpA[OmpA-linker] Cr |
29 Electrophoresis of OmpA[J23101+B0034+OmpA+pSB3C5] Transform of GBP[J23110+B0034+FadL-GBP+B0030+E0040+J61048+pSB1C3] Cr GBP[R0010+B0034+FadL-GBP+B0030+E0040+J661048+pSB6A1] Ar PCR of BFP[B0034+K592100+J61048+pSB1C3] Cr GBP[R0010+B0034+FadL-GBP+B0030+E0040+J61048] Electrophoresis of BFP[B0034+K592100+J61048+pSB1C3] Cr GBP[R0010+B0034+FadL-GBP+B0030+E0040+J61048] Digestion of Anti-Her 2[Herceptin-own] (EcoRI and PstI) Anti-Her 2[Herceptin-E1010] (EcoRI and PstI) GBP[GBP] (EcoRI and PstI) Ligation of [R0010+B0034+pSB1K3] Kr BFP[B0030+K592100+J61048] Cr GBP[R0010+B0034+FadL-GBP+B0034+E0040+J61048] Ar GFP[R0010+B0030+E0040+J61048] Ar Electrophoresis of Anti-Her 2[Herceptin-own] (EcoRI and PstI) Anti-Her 2[Herceptin-E1010] (EcoRI and PstI) GBP[GBP] (EcoRI and PstI) [R0010+B0034+pSB1K3] Kr BFP[B0030+K592100+J61048] Cr GBP[R0010+B0034+FadL-GBP+B0034+E0040+J61048] Ar GFP[R0010+B0030+E0040+J61048] Ar |
30 Cultivation of scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB1C3] Cr scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB1C3] Cr Ligation of GBP[FadL-GBP+pSB1C3]Cr Digestion of Anti-Her 2[Herceptin own] (NcoI and PstI) [pSB1A3] (EcoRI and PstI) [pSB1C3] (EcoRI and PstI) Cultivation of scFv-C[J23101+B0034+OmpA+scFv-C] Ar [J23101+B0034] Kr Digestion of scFv-B[scFv-B] Kr (NcoI and PstI) scFv-C[scFv-C] Kr (NcoI and PstI) OmpA[OmpA] Cr (EcoRI and PstI) Electrophoresis of Anti-Her 2[Herceptin own] (NcoI and PstI) [pSB1A3] (EcoRI and PstI) [pSB1C3] (EcoRI and PstI) Plasmid purification of scFv-C[J23101+B0034+OmpA+scFv-C] Ar [J231-1+B0034] Kr Digestion of scFv-C[J23101+B0034+OmpA+scFv-C] Ar (EcoRI and SpeI) [J231-1+B0034] Kr (EcoRI and SpeI) Anti-Her 2[Herceptin-Own] (NcoI and PstI) |
31 Electrophoresis of scFv-C[J23101+B0034+OmpA+scFv-C] Ar (EcoRI and SpeI) [J231-1+B0034] Kr (EcoRI and SpeI) Anti-Her 2[Herceptin-Own] (NcoI and PstI) Ligation of Anti-Her 2[J2310+B0034+OmpA+Herceptin-Own+pSB1A3] Ar Transform of Anti-Her 2[J2310+B0034+OmpA+Herceptin-Own+pSB1A3] Ar Electrophoresis of [pSB1C3] (XbaI and PstI) OmpA[OmpA] Cr (EcoRI and PstI) scFv-B[scFv-B] Kr (NcoI and PstI) scFv-C[scFv-C] Kr (NcoI and PstI) |
| Sun. | Mon. | Tue. | Wed. | Thu. | Fri. | Set. |
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1 Digestion of Anti-Her 2[Herceptin-Web] (NcoI and PstI) Electrophoresis of Anti-Her 2[Herceptin-Web] (NcoI and PstI) | ||||||
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2 Electrophoresis of [pSB1C3] (XbaI and PstI) OmpA[OmpA] (EcoRI and PstI) scFv-B[scFv-B] Kr (NcoI and PstI) scFv-C[scFv-C] Kr (NcoI and PstI) OmpA[OmpA] (NcoI and PstI) Digestion of BFP[OmpA+K592100] Cr (EcoRI and NcoI) |
3 Transform of [R0010+B0034+pSB1K3] Kr GBP[FadL-GBP+pSB1C3] Cr BFP[B0030+K592100+J61048] Cr Digestion of Anti-Her 2[Herceptin-Web] (NcoI and PstI) Ligation of Anti-Her 2[J23101+B0034+OmpA+Herceptin-Web] Ar |
4 Digestion of BFP[OmpA+K592100] Cr (EcoRI and NcoI) PCR of [R0010+B0034+pSB1K3] Kr BFP[B0030+K592100+J61048] Cr Transform of Anti-Her 2[J23101+B0034+OmpA+Herceptin-Web] Ar |
5 Digestion of [pSB6A1] (EcoRI and PstI) [pSB1C3] (EcoRI and PstI) PCR of Anti-Her 2[J23101+B0034+OmpA+Herceptin-Web] Ar Electrophoresis of Anti-Her 2[J23101+B0034+OmpA+Herceptin-Web] Ar Cultivation of BFP[B0030+K592100+J61048] Cr GBP[R0010+B0034+FadL-GBP+B0030+E0040+J61048] PCR of GBP[FadL-GBP] Cr BFP[OmpA+K592100] Cr scFv-C[J23101+B0034+OmpA+scFv-C] Ar scFv-C[scFv-C+pSB1C3] Cr scFv-B[scFv-B+oSB1C3] Cr Digestion of scFv-C[J23101+B0034+OmpA+scFv-C+pSB3C5] Cr (EcoRI and PstI) scFv-B[J23101+B0034+OmpA+scFv-B+pSB3C5] Cr (EcoRI and PstI) Transform of GBP[J23101+B0034+FadL-GBP]Cr scFv-B[scFv-B+pSB1C3] Cr scFv-C[scFv-C+pSB1C3] Cr scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048] Cr Gel extraction of [pSB6A1] (EcoRI and PstI) [OmpA] (NcoI and PstI) [pSB1C3] (EcoRI and PstI) |
6 Cultivation of Anti-Her 2[J23101+B0034+OmpA+Herceptin-Web] Ar Ligation of Anti-Her 2[OmpA+Herceptin-Web] Cr Anti-Her 2[OmpA+Herceptin-Own] Cr scFv-C[OmpA+scFv-C] Cr scFv-B[OmpA+scFv-B] Cr scFv-C[J23101+B0034+OmpA+scFv-C] Cr scFv-B[J23101+B0034+OmpA+scFv-B] Cr Transform of Anti-Her 2[OmpA+Herceptin-Web] Cr Anti-Her 2[OmpA+Herceptin-Own] Cr scFv-C[OmpA+scFv-C] Cr scFv-B[OmpA+scFv-B] Cr scFv-C[J23101+B0034+OmpA+scFv-C] Cr scFv-B[J23101+B0034+OmpA+scFv-B] Cr [pSB1C3] [pSB1A3] [pSB1K3] Plasmid purification of BFP[B0030+K592100+J61048] Digestion of BFP[B0030+K592100+J61048] (EcoRI and PstI) Cultivation of GBP[R0010+B0034+FadL-GBP+B0030+E0040+J61048] Cr Ligation of [R0010+B0034] Kr GBP[GBP[J23101+B0034+FadL-GBP]+pSB1C3] Cr Digestion of [pSB1C3] (XbaI and PstI) Electrophoresis of GBP[J23101+B0034+FadL-GBP] Cr scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040] Cr scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040] Cr scFv-C[scFv-C] Cr scFv-B[scFv-B] Cr Plasmid purification of Anti-Her 2[J23101+B0034+OmpA+Herceptin-Web] Ar Digestion of Anti-Her 2[J23101+B0034+OmpA+Herceptin-Web] Ar (EcoRI and PstI) Ligation of Anti-Her 2[J23101+B0034+OmpA+Herceptin-Own] Ar Electrophoresis of Anti-Her 2[J23101+B0034+OmpA+Herceptin-Web] Ar Anti-Her 2[J23101+B0034+OmpA+Herceptin-Own] Ar [B0034] [pSB1C3] Cultivation of Anti-Her 2[J23101+B0034+OmpA+Herceptin-Web] Ar Transform of GBP[J23101+B0034+FadL-GBP] Ar Electrophoresis of scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040] Cr scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040] Cr [pSB1C3] (XbaI and PstI) [pSB6A1] (XbaI and PstI) [pSB1C3] (XbaI and PstI) Anti-Her 2[J23101+B0034+OmpA+Herceptin-Web] Ar Cultivation of scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048] Cr scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048] Cr GFP[J23101+B0034+E0040+J61048] Ar SDS-page of GFP[J23101+B0034+E0040+J61048] Ar scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048] Ar scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048] Ar [pSB6A1] GBP[J23101+B0034+FadL-GBP+B0030+E0040+J61048] Ar Digestion of [pSB1C3] (XbaI and PstI) Anti-Her 2[Herceptin-Web] (NcoI and PstI) Anti-Her 2[Herceptin-Own] (NcoI and PstI) Ligation of [pSB1C3] (XbaI and PstI) Anti-Her 2[Herceptin-Web] Cr Anti-Her 2[Herceptin-Own] Cr BFP[J23101+B0034+OmpA+K592100] Cr |
7 Plasmid purification of GBP[R0010+B0034+FadL-GBP+B0030+E0040+J61048] Cr Digestion of GBP[R0010+B0034+FadL-GBP+B0030+E0040+J61048] Cr PCR of scFv-B[J23101+B0034+OmpA+scFv-B] Cr scFv-C[J23101+B0034+OmpA+scFv-C] Cr Electrophoresis of Anti-Her 2[J23101+B0034+OmpA+Herceptin-Web] Ar (EcoRI and PstI) Cultivation of scFv-B[J23101+B0034+OmpA+scFv-B] Cr scFv-C[J23101+B0034+OmpA+scFv-C] Cr Transform of Anti-Her 2[J23101+B0034+OmpA+Herceptin-Web] Ar Plasmid purification of GBP[J23101+B0034+FadL-GBP+B0030+E0040+J61048] Cr Digestion of GBP[J23101+B0034+FadL-GBP+B0030+E0040+J61048] Cr (EcoRI and PstI) PCR of Anti-Her 2[J23101+B0034+OmpA+Herceptin-Own] Ar Electrophoresis of scFv-B[J23101+B0034+OmpA+scFv-B] Cr scFv-C[J23101+B0034+OmpA+scFv-C] Cr Anti-Her 2[J23101+B0034+OmpA+Herceptin-Own] Ar OmpA[OmpA] (NcoI and PstI) [pSB1C3] (EcoRI and PstI) Ligation of scFv-C[OmpA+scFv-C] Cr scFv-B[OmpA+scFv-B] Cr Anti-Her 2[Herceptin-Own] Cr Anti-Her 2[Herceptin-Web] Cr Anti-Her 2[OmpA+Herceptin-Own] Cr Anti-Her 2[OmpA+Herceptin-Web] C Electrophoresis of Anti-Her 2[J23101+B0034+OmpA+Herceptin-Web] Ar (EcoRI and PstI) Anti-Her 2[Herceptin-Web] (NcoI and PstI) Anti-Her 2[Herceptin-Own] (NcoI and PstI) Transform of scFv-C[OmpA+scFv-C] Cr scFv-B[OmpA+scFv-B] Cr Anti-Her 2[Herceptin-Own] Cr Anti-Her 2[Herceptin-Web] Cr Anti-Her 2[OmpA+Herceptin-Own] Cr Anti-Her 2[OmpA+Herceptin-Web] C BFP[J23101+B0034+OmpA+K592100] Cr |
8 Plasmid purification of scFv-C[J23101+OmpA+scFv-C] Cr scFv-B[J23101+OmpA+scFv-B] Cr Digestion of scFv-C[J23101+OmpA+scFv-C] Cr (EcoRI and PstI) scFv-B[J23101+OmpA+scFv-B] Cr (EcoRI and PstI) PCR of Anti-Her 2[J23101+B0034+OmpA+Herceptin-Own] Ar Cultivation of [pSB1K3] [pSB1C3] [pSB1A3] PCR of BFP[J23101+B0034+OmpA+K592100] Cr Transform of scFv-C[OmpA+scFv-C] Cr Electrophoresis of BFP[J23101+B0034+OmpA+K592100] Cr scFv-C[J23101+OmpA+scFv-C] Cr (EcoRI and PstI) scFv-B[J23101+OmpA+scFv-B] Cr (EcoRI and PstI) [pSB1C3] (EcoRI and PstI) Digestion of Anti-Her 2[J23101+B0034+OmpA+Herceptin-Web] Ar (EcoRI and SpeI) OmpA[OmpA] Cr NcoI and PstI OmpA[OmpA+K592100] Cr NcoI and PstI Ligation of Anti-Her 2[J23101+B0034+OmpA+Herceptin-Web] Cr Digestion of Anti-Her 2[Herceptin-Own] (EcoRI and PstI) Anti-Her 2[Herceptin-Web] (EcoRI and PstI) Electrophoresis of BFP[J23101+B0034+OmpA+K592100] Cr |
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9 Cultivation of Anti-Her 2[J23101+B0034+OmpA+Herceptin-Own] Ar Plasmid purification of [pSB1C3] [pSB1K3] [pSB1A3] Digestion of [pSB1C3] (EcoRI and PstI) [pSB1K3] (EcoRI and PstI) [pSB1A3] (EcoRI and PstI) Transform of Anti-Her 2[Herceptin-Own] Cr Anti-Her 2[Herceptin-Web] Cr Anti-Her 2[J23101+B0034+OmpA+Herceptin-Web] Cr [R0010+B0034] Cr BFP[J23101+B0034+OmpA+K592100] Cr Electrophoresis of Anti-Her 2[J23101+B0034+OmpA+Herceptin-Web] Cr (EcoRI and SpeI) OmpA[OmpA] Cr (NcoI and PstI) Digestion of [pSB1A3] (EcoRI and PstI) [pSB1K3] (EcoRI and PstI) [pSB1C3] (EcoRI and PstI) RFP[B0030+E1010+J61048] Cr (XbaI and PstI) Electrophoresis of scFv-C[OmpA+scFv-C] Kr PCR of GBP[GBp+pSB1C3] Cr Digestion of BFP[B0030+K592100+J61048] Cr (XbaI and PstI) |
10 Electrophoresis of GBP[GBp+pSB1C3] Cr [pSB1A3] (EcoRI and PstI) [pSB1K3] (EcoRI and PstI) [pSB1C3] (EcoRI and PstI) Ligation of Anti-Her 2[J23101+B0034+OmpA+Herceptin-Web+B0030+E1010+J61048] Kr Cultivation of [R0010+B0034] Cr BFP[B0030+K592100+J61048] Cr scFv-C[OmpA+scFv-C] Kr Digestion of [pSB1C3] (EcoRI and Pstl) Transform of scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB6A1] Ar scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB6A1] Ar GBP[R0010+B0034+FadL-GBP+B0030+E0040+J61048+pSB1K3] Kr Gel extraction of [pSB1C3] (EcoRI and PstI) PCR of Anti-Her 2[J23101+B0034+OmpA+Herceptin-Web] Anti-Her 2[Herceptin-Web] Anti-Her 2[Herceptin-Own] |
11 Plasmid purification of [R0010+B0034] Cr scFv-C[OmpA+scFv-C] Kr BFP[OmpA+K592100] Cr Gel extraction of [pSB1C3] (EcoRI and PstI) Digestion of [R0010+B0034] Cr (EcoRI and SpeI) scFv-C[OmpA+scFv-C] Kr (EcoRI and PstI) BFP[OmpA+K592100] Cr (EcoRI and PstI) GBP[J23101+B0034+FadL-GBP] Cr (EcoRI and SpeI) Ligation of Anti-Her 2[Herceptin-Web] Cr PCR of scFv-B[OmpA+scFv-B] Kr Electrophoresis of [R0010+B0034] Cr (EcoRI and SpeI) scFv-C[OmpA+scFv-C] Kr (EcoRI and PstI) BFP[OmpA+K592100] Cr (EcoRI and PstI) GBP[J23101+B0034+FadL-GBP] Cr (EcoRI and SpeI) Ligation of GBP[J23101+B0034+FadL-GBP+B0030+E0040+J61048+pSB6A1] Ar Electrophoresis of scFv-B[OmpA+scFv-B] Kr PCR of scFv-C[J23101+B0034+OmpA+scFv-B+B0030+K592100+J61048] Kr Transform of GBP[R0010+B0034+FadL-GBP+B0030+E0040+J61048] Kr |
12 Electrophoresis of scFv-B[OmpA+scFv-B] Kr Transform of Anti-Her 2[Herceptin-Web] Cr Anti-Her 2[Herceptin-Own] Cr Anti-Her 2[J23101+B0034+OmpA+Herceptin-Web] Ar PCR of scFv-C[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB6A1] Ar Digestion of [pSB1C3] (EcoRI and PstI) Electrophoresis of scFv-C[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB6A1] Ar |
13 Transform of GBP[J231010+B0034+FadL-GBP+B0030+E0040+J61048+pSB6A1] Ar Cultivation of [pSB1C3] scFv-B[OmpA-scFv-B] Kr Plasmid purification of [pSB1C3] scFv-B[OmpA-scFv-B] Kr Digestion of [pSB1C3] (EcoRI and PstI) scFv-B[OmpA-scFv-B] Kr (EcoRI and PstI) Electrophoresis of [pSB1C3] (EcoRI and PstI) Ligation of OmpA[R0010+B0034+OmpA+pSB1A3] Ar GBP[R0010+B0034+FadL-GBP+pSB1A3] Ar scFv-B[J23101+B0034+scFv-B+B0030+E0040+J61048+pSB6A1] Ar scFv-C[J23101+B0034+scFv-C+B0030+E0040+J61048+pSB6A1] Ar Electrophoresis of [pSB1C3] (EcoRI and PstI) scFv-B[OmpA-scFv-B] Kr (EcoRI and PstI) |
14 Transform of OmpA[R0010+B0034+OmpA+pSB1A3] Ar GBP[R0010+B0034+FadL-GBP+pSB1A3] Ar scFv-C[J23101+B0034+scFv-C+B0030+E0040+J61048+pSB6A1] Ar |
15 PCR of OmpA[R0010+B0034+OmpA+pSB1A3] Ar GBP[R0010+B0034+FadL-GBP+pSB1A3] Ar scFv-C[J23101+B0034+scFv-C+B0030+E0040+J61048+pSB6A1] Ar Ligation of scFv-C[J23101+B0034+OmpA+scFv-C+pSB3C5] Cr BFP[J23101+B0034+OmpA+K592100+pSB3C5] Cr PCR of GBP[R0010+B0034+FadL-GBP+pSB1A3] Kr GBP[R0010+B0034+FadL-GBP+B0030+E0040+J61048] Kr Transform of scFv-B[J23101+B0034+OmpA+scFv-B+B0030+K592100+J61048] Kr Electrophoresis of GBP[R0010+B0034+FadL-GBP+pSB1A3] Kr GBP[R0010+B0034+FadL-GBP+B0030+E0040+J61048] Kr Electrophoresis of GBP[FadL-GBP] (EcoRI and PstI) |
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16 PCR of GBP[R0010+B0034+FadL-GBP+pSB1A3] Kr GBP[R0010+B0034+FadL-GBP+B0030+E0040+J61048] Kr scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048] Kr Cultivation of OmpA[R0010+B0034+OmpA] Electrophoresis of GBP[R0010+B0034+FadL-GBP+pSB1A3] Kr GBP[R0010+B0034+FadL-GBP+B0030+E0040+J61048] Kr scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048] Kr |
17 Ligation of scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB6A1] Ar scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB6A1] Ar Transform of scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB6A1] Ar scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB6A1] Ar |
18 Plasmid purification of OmpA[R0010+B0034+OmpA+pSB1A3] Ar GBP[R0010+B0034+FadL-GBP+pSB1A3] Ar Digestion of BFP[OmpA+K592100] Cr (EcoRI and NcoI) Anti-Her 2[Herceptin-Own] (NcoI and SpeI) Anti-Her 2[Herceptin-Web] (NcoI and SpeI) Anti-Her 2[J23101+B0034+OmpA+Herceptin-Own] Ar (EcoRI and SpeI) BFP[B0030+K592100+J61048] Cr (XbaI and PstI) OmpA[R0010+B0034+OmpA+pSB1A3] Ar (EcoRI and PstI) GBP[R0010+B0034+FadL-GBP+pSB1A3] Ar (EcoRI and PstI) Ligation of Anti-Her 2[J23101+B0034+OmpA+Herceptin-Own+pSB1C3] Cr GBP[R0010+B0034+FadL-GBP+pSB1A3] Ar Transform of Anti-Her 2[J23101+B0034+OmpA+Herceptin-Own+pSB1C3] Cr GBP[R0010+B0034+FadL-GBP+pSB1A3] Ar GBP[J23101+B0034+fadL-GBP+B0030+E0040+J61048] Cr OmpA[R0010+B0034+OmpA+pSB1C3] Cr GBP[R0010+B0034+FadL-GBP+pSB1C3] Cr scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048] Ligation of scFv-B[R0010+B0034+OmpA+scFv-B] scFv-C[R0010+B0034+OmpA+scFv-C] Anti-Her 2[R0010+B0034+OmpA+Herceptin-Own] Anti-Her 2[R0010+B0034+OmpA+Herceptin-Web] GBP[R0010+B0034+FadL-GBP+B0030+E0040+J61048] PCR of Anti-Her 2[Herceptin-Web] scFv-B[J23101+B0034+OmpA+scFv-B] scFv-C[J23101+B0034+OmpA+scFv-C+pSB3C5] Cr Anti-Her 2[Herceptin-Own] scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB6A1] Ar |
19 Electrophoresis of Anti-Her 2[Herceptin-Web] scFv-B[J23101+B0034+OmpA+scFv-B] scFv-C[J23101+B0034+OmpA+scFv-C+pSB3C5] Cr Anti-Her 2[Herceptin-Own] scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB6A1] Ar scFv-C[OmpA+scFv-C] Anti-Her 2[Herceptin-Own] (NcoI and SpeI) Anti-Her 2[Herceptin-Web] (NcoI and SpeI) Anti-Her 2[J23101+B0034+OmpA+Herceptin-Own] Ar (EcoRI and SpeI) BFP[B0030+K592100+J61048] Cr (XbaI and PstI) Transform of GBP[R0010+B034+FadL-GBP] Ar GBP[J23101+B0034+FadL-GBP] Ar GBP[J23101+B0034+FadL-GBP+B0030+E0040+J61048] Cr |
20 PCR of GBP[FadL-GBP] Kr OmpA[OmpA] Cr Gel extraction of OmpA[OmpA] (EcoRI and SpeI) Plasmid purification of Anti-Her 2[Herceptin-Own] Cr scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB6A1] Ar PCR of GBP[FadL-GBP] Kr OmpA[R0010+B0034+OmpA+pSB1C3] Cr GBP[R0010+B0034+GBP+pSB1C3] Cr scFv-B[J23101+B0034+OmpA+scFv-B+B0030+K592100+J61048] Kr GBP[R0010+B0034+GBP+B0030+E0040+J61048] scFv-B[R0010+B0034+OmpA+scFv-B] scFv-C[R0010+B0034+OmpA+scFv-C] Anti-Her 2[R0010+B0034+OmpA+Herceptin-Own] Anti-Her 2[R0010+B0034+OmpA+Herceptin-Web] Digestion of scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB6A1] Ar (EcoRI and PstI) Anti-Her 2[Herceptin-Own] Cr Electrophoresis of OmpA[R0010+B0034+OmpA+pSB1C3] Cr GBP[R0010+B0034+GBP+pSB1C3] Cr scFv-B[J23101+B0034+OmpA+scFv-B+B0030+K592100+J61048] Kr GBP[R0010+B0034+GBP+B0030+E0040+J61048] Ligation of scFv-B[R0010+B0034+OmpA+scFv-B+pSB1K3] Kr scFv-C[R0010+B0034+OmpA+scFv-C+pSB1K3] Kr Anti-Her 2[R0010+B0034+OmpA+Herceptin-Own+pSB1K3] Kr Anti-Her 2[R0010+B0034+OmpA+Herceptin-Web+pSB1K3] Kr GBP[R0010+B0034+GBP+B0030+E0040+J61048pSB1A3] Ar scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E1010+J61048+pSB1K3] Kr scFv-B[J23101+B0034+OmpA+scFv-B+B0030+K592100+J61048+pSB1K3] Kr Anti-Her 2[R0010+B0034+OmpA+Herceptin-Own+B0030+E1010+J61048+pSB1K3] Kr |
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