Difference between revisions of "Team:NCTU Formosa/Notebook"
| Line 559: | Line 559: | ||
<td > | <td > | ||
<B>15</B><br><br> | <B>15</B><br><br> | ||
| − | Transformation of GBP[FadL-GBP]<br> | + | Transformation of <br>GBP[FadL-GBP]<br> |
| − | Cultivation | + | Cultivation of <br>OmpA[OmpA+pSB1K3] K<sup>r</sup> |
</td> | </td> | ||
<td> | <td> | ||
<B>16</B><br><br> | <B>16</B><br><br> | ||
| − | Cultivation of GBP[FadL-GBP] | + | Cultivation of <br>GBP[FadL-GBP] |
<br> | <br> | ||
| − | Plasmid purification of OmpA[OmpA+pSB1K3] K<sup>r</sup> | + | Plasmid purification of <br>OmpA[OmpA+pSB1K3] K<sup>r</sup> |
<br> | <br> | ||
| − | Digestion of OmpA[OmpA+pSB1K3] K<sup>r</sup> (EcoRI and PstI) | + | Digestion of <br>OmpA[OmpA+pSB1K3] K<sup>r</sup> (EcoRI and PstI) |
</td> | </td> | ||
<td> | <td> | ||
<B>17</B><br><br>Plasmid purification of | <B>17</B><br><br>Plasmid purification of | ||
| − | GBP[FadL-GBP] | + | <br>GBP[FadL-GBP] |
<br> | <br> | ||
Digestion of | Digestion of | ||
| − | GBP[FadL-GBP] (EcoRI and PstI) | + | <br>GBP[FadL-GBP] (EcoRI and PstI) |
<br> | <br> | ||
Electrophoresis of digested product: | Electrophoresis of digested product: | ||
| − | GBP[FadL-GBP] (EcoRI and PstI) | + | <br>GBP[FadL-GBP] (EcoRI and PstI) |
</td> | </td> | ||
| Line 585: | Line 585: | ||
<B>18</B><br><br> | <B>18</B><br><br> | ||
Transformation of ligated product: | Transformation of ligated product: | ||
| − | GBP[FadL-GBP+pSB1C3] C<sup>r</sup> | + | <br>GBP[FadL-GBP+pSB1C3] C<sup>r</sup> |
</td> | </td> | ||
| Line 593: | Line 593: | ||
<B>19</B><br><br> | <B>19</B><br><br> | ||
Cultivation of | Cultivation of | ||
| − | ligated product: GBP[FadL-GBP] C<sup>r</sup> | + | <br>ligated product: GBP[FadL-GBP] C<sup>r</sup> |
| − | PCR product: Terminator[J61048] | + | <br>PCR product: Terminator[J61048] |
</td> | </td> | ||
| Line 603: | Line 603: | ||
<B>21</B><br><br> | <B>21</B><br><br> | ||
Cultivation of | Cultivation of | ||
| − | GFP[J23101+B0034+E0040+J61048+pSB1A3] A<sup>r</sup> | + | <br>GFP[J23101+B0034+E0040+J61048+pSB1A3] A<sup>r</sup> |
| − | OmpA[OmpA+pSB1C3] C<sup>r</sup> | + | <br>OmpA[OmpA+pSB1C3] C<sup>r</sup> |
| − | GBP[FadL-GBP+pSB1C3] C<sup>r</sup> | + | <br>GBP[FadL-GBP+pSB1C3] C<sup>r</sup> |
</td> | </td> | ||
| Line 611: | Line 611: | ||
<B>22</B><br><br> | <B>22</B><br><br> | ||
Plasmid purification of | Plasmid purification of | ||
| − | GFP[J23101+B0034+E0040+J61048+pSB1A3] A<sup>r</sup> | + | <br>GFP[J23101+B0034+E0040+J61048+pSB1A3] A<sup>r</sup> |
| − | OmpA[OmpA+pSB1C3] C<sup>r</sup> | + | <br>OmpA[OmpA+pSB1C3] C<sup>r</sup> |
| − | GBP[FadL-GBP+PSB1C3] C<sup>r</sup> | + | <br>GBP[FadL-GBP+PSB1C3] C<sup>r</sup> |
| − | + | <br> | |
Digestion of | Digestion of | ||
| − | GFP[J23101+B0034+E0040+J61048] A<sup>r</sup> (EcoRI and PstI) | + | <br>GFP[J23101+B0034+E0040+J61048] A<sup>r</sup> (EcoRI and PstI) |
| − | OmpA[OmpA] C<sup>r</sup> (EcoRI and PstI) | + | <br>OmpA[OmpA] C<sup>r</sup> (EcoRI and PstI) |
| − | GBP[FadL-GBP] C<sup>r</sup> (EcoRI and PstI) | + | <br>GBP[FadL-GBP] C<sup>r</sup> (EcoRI and PstI) |
| − | + | <br> | |
Electrophoresis of digested product: | Electrophoresis of digested product: | ||
| − | GFP[J23101+B0034+E0040+J61048] A<sup>r</sup> (EcoRI and PstI) | + | <br>GFP[J23101+B0034+E0040+J61048] A<sup>r</sup> (EcoRI and PstI) |
| − | OmpA[OmpA] C<sup>r</sup> (EcoRI and PstI) | + | <br>OmpA[OmpA] C<sup>r</sup> (EcoRI and PstI) |
| − | GBP[FadL-GBP] C<sup>r</sup> (EcoRI and PstI) | + | <br>GBP[FadL-GBP] C<sup>r</sup> (EcoRI and PstI) |
</td> | </td> | ||
| Line 630: | Line 630: | ||
Digestion of | Digestion of | ||
OmpA[OmpA] C<sup>r</sup> (EcoRI and SpeI) | OmpA[OmpA] C<sup>r</sup> (EcoRI and SpeI) | ||
| − | GBP[FadL-GBP] C<sup>r</sup> (EcoRI and SpeI) | + | <br>GBP[FadL-GBP] C<sup>r</sup> (EcoRI and SpeI) |
| − | + | <br> | |
Electrophoresis of digested product: | Electrophoresis of digested product: | ||
| − | OmpA[OmpA] C<sup>r</sup> (EcoRI and SpeI) | + | <br>OmpA[OmpA] C<sup>r</sup> (EcoRI and SpeI) |
| − | GBP[FadL-GBP] C<sup>r</sup> (EcoRI and SpeI) | + | <br>GBP[FadL-GBP] C<sup>r</sup> (EcoRI and SpeI) |
</td> | </td> | ||
| Line 640: | Line 640: | ||
<B>24</B><br><br> | <B>24</B><br><br> | ||
Digestion of | Digestion of | ||
| − | OmpA[OmpA] C<sup>r</sup> (XbaI and PstI) | + | <br>OmpA[OmpA] C<sup>r</sup> (XbaI and PstI) |
| − | GBP[FadL-GBP] C<sup>r</sup> (XbaI and PstI) | + | <br>GBP[FadL-GBP] C<sup>r</sup> (XbaI and PstI) |
| Line 648: | Line 648: | ||
<B>25</B><br><br> | <B>25</B><br><br> | ||
Electrophoresis of digested product: | Electrophoresis of digested product: | ||
| − | OmpA[OmpA] C<sup>r</sup> (XbaI and PstI) | + | <br>OmpA[OmpA] C<sup>r</sup> (XbaI and PstI) |
| − | GBP[FadL-GBP] C<sup>r</sup> (XbaI and PstI) | + | <br>GBP[FadL-GBP] C<sup>r</sup> (XbaI and PstI) |
| − | + | <br> | |
Digestion of | Digestion of | ||
| − | RBS[B0034] A<sup>r</sup> (XbaI and PstI) | + | <br>RBS[B0034] A<sup>r</sup> (XbaI and PstI) |
| − | backbone[pSB1A3] A<sup>r</sup> (EcoRI and PstI) | + | <br>backbone[pSB1A3] A<sup>r</sup> (EcoRI and PstI) |
| − | Terminator[J61048] A<sup>r</sup> (XbaI and PstI) | + | <br>Terminator[J61048] A<sup>r</sup> (XbaI and PstI) |
| − | + | <br> | |
Electrophoresis of digested product: | Electrophoresis of digested product: | ||
| − | RBS[B0034] A<sup>r</sup> (XbaI and PstI) | + | <br>RBS[B0034] A<sup>r</sup> (XbaI and PstI) |
| − | backbone[pSB1A3] A<sup>r</sup> (EcoRI and PstI) | + | <br>backbone[pSB1A3] A<sup>r</sup> (EcoRI and PstI) |
| − | Terminator[J61048] A<sup>r</sup> (XbaI and PstI) | + | <br>Terminator[J61048] A<sup>r</sup> (XbaI and PstI) |
| − | + | <br> | |
Ligation of | Ligation of | ||
| − | OmpA[J23101+B0034+OmpA+pSB1A3] A<sup>r</sup> | + | <br>OmpA[J23101+B0034+OmpA+pSB1A3] A<sup>r</sup> |
| − | GBP[GBP+B0034+pSB1K3] K<sup>r</sup> | + | <br>GBP[GBP+B0034+pSB1K3] K<sup>r</sup> |
| − | OmpA[OmpA+J61048+pSB1K3] K<sup>r</sup> | + | <br>OmpA[OmpA+J61048+pSB1K3] K<sup>r</sup> |
</td> | </td> | ||
| Line 672: | Line 672: | ||
<B>26</B><br><br> | <B>26</B><br><br> | ||
Ligation of | Ligation of | ||
| − | RFP[E1010+B0034+pSB1K3] K<sup>r</sup> | + | <br>RFP[E1010+B0034+pSB1K3] K<sup>r</sup> |
| − | + | <br> | |
Transformation of ligated product: | Transformation of ligated product: | ||
| − | OmpA[J23101+B0034+OmpA+pSB1A3] A<sup>r</sup> | + | <br>OmpA[J23101+B0034+OmpA+pSB1A3] A<sup>r</sup> |
| − | GBP[FadL-GBP+B0034+pSB1K3] K<sup>r</sup> | + | <br>GBP[FadL-GBP+B0034+pSB1K3] K<sup>r</sup> |
| − | OmpA[OmpA+J61048+pSB1K3] K<sup>r</sup> | + | <br>OmpA[OmpA+J61048+pSB1K3] K<sup>r</sup> |
| − | + | <br> | |
Electrophoresis of digested product: | Electrophoresis of digested product: | ||
| − | RFP[E1010+J61048+pSB1K3] K<sup>r</sup> | + | <br>RFP[E1010+J61048+pSB1K3] K<sup>r</sup> |
| − | [J23101+B0034] | + | <br>[J23101+B0034] |
</td> | </td> | ||
| Line 687: | Line 687: | ||
<B>27</B><br><br> | <B>27</B><br><br> | ||
Digestion of | Digestion of | ||
| − | Terminator[J61048] A<sup>r</sup> (XbaI and PstI) | + | <br>Terminator[J61048] A<sup>r</sup> (XbaI and PstI) |
| − | + | <br> | |
| − | PCR of ligated product | + | PCR of ligated product: |
| − | OmpA[J23101+B0034+OmpA+pSB1A3] A<sup>r</sup> | + | <br>OmpA[J23101+B0034+OmpA+pSB1A3] A<sup>r</sup> |
| − | OmpA[OmpA+J61048+pSB1K3] K<sup>r</sup> | + | <br>OmpA[OmpA+J61048+pSB1K3] K<sup>r</sup> |
</td> | </td> | ||
| Line 697: | Line 697: | ||
<B>28</B><br><br> | <B>28</B><br><br> | ||
Electrophoresis of | Electrophoresis of | ||
| − | RFP[E1010+J61048+pSB1K3] K<sup>r</sup> | + | <br>RFP[E1010+J61048+pSB1K3] K<sup>r</sup> |
| − | OmpA[J23101+B0034+OmpA+pSB1A3] A<sup>r</sup> | + | <br>OmpA[J23101+B0034+OmpA+pSB1A3] A<sup>r</sup> |
Transformation of ligated product: | Transformation of ligated product: | ||
| − | GBP[B0034+FadL-GBP+pSB1A3] A<sup>r</sup> | + | <br>GBP[B0034+FadL-GBP+pSB1A3] A<sup>r</sup> |
| − | OmpA[OmpA+J61048+pSB1K3] K<sup>r</sup> | + | <br>OmpA[OmpA+J61048+pSB1K3] K<sup>r</sup> |
PCR of ligated product | PCR of ligated product | ||
| − | OmpA[J23101+B0034+OmpA] A<sup>r</sup> | + | <br>OmpA[J23101+B0034+OmpA] A<sup>r</sup> |
</td> | </td> | ||
| Line 711: | Line 711: | ||
<B>29</B><br><br> | <B>29</B><br><br> | ||
Cultivation of | Cultivation of | ||
| − | RFP[E1010+J61048+pSB1K3] K<sup>r</sup> | + | <br>RFP[E1010+J61048+pSB1K3] K<sup>r</sup> |
PCR of | PCR of | ||
| − | OmpA[J23101+B0034+OmpA] A<sup>r</sup> | + | <br>OmpA[J23101+B0034+OmpA] A<sup>r</sup> |
| − | Electrophoresis of | + | <br>Electrophoresis of |
| − | OmpA[J23101+B0034+OmpA] A<sup>r</sup> | + | <br>OmpA[J23101+B0034+OmpA] A<sup>r</sup> |
Ligation of | Ligation of | ||
| − | GBP[FadL-GBP+J61048+pSB1K3] K<sup>r</sup> | + | <br>GBP[FadL-GBP+J61048+pSB1K3] K<sup>r</sup> |
</td> | </td> | ||
| Line 727: | Line 727: | ||
<B>30</B><br><br> | <B>30</B><br><br> | ||
Plasmid purification of | Plasmid purification of | ||
| − | RFP[E1010+J61048+pSB1K3] K<sup>r</sup> | + | <br>RFP[E1010+J61048+pSB1K3] K<sup>r</sup> |
Ligation of | Ligation of | ||
| − | OmpA[J23101+B0034+OmpA] A<sup>r</sup> | + | <br>OmpA[J23101+B0034+OmpA] A<sup>r</sup> |
PCR of | PCR of | ||
| − | OmpA[J23101+B0034+OmpA] A<sup>r</sup> | + | <br>OmpA[J23101+B0034+OmpA] A<sup>r</sup> |
Transformation of | Transformation of | ||
| − | 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> |
</td> | </td> | ||
Revision as of 07:38, 11 September 2015
Lab note
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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 |
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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 |
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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 |
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