Difference between revisions of "Team:CityU HK/Notebook/Lysis plasmid (λ phage)"
Tommykong2 (Talk | contribs) |
|||
Line 346: | Line 346: | ||
<b>co</b>: codon optimized <br> | <b>co</b>: codon optimized <br> | ||
− | <b>λ Lysis(o_o_o)</b>: SλWT(ori)-Rλ(ori)-Rzλ(ori) <br> | + | <b>λ Lysis(o_o_o)</b>: <i>SλWT(ori)-Rλ(ori)-Rzλ(ori)</i> <br> |
− | <b>λLysis(c_c_c)</b>: SλWT(co)-Rλ(co)-Rzλ(co) <br> | + | <b>λLysis(c_c_c)</b>: <i>SλWT(co)-Rλ(co)-Rzλ(co) <br></i> |
− | <b>λLysis_Sm(o_c_c)</b>: Sλmut(ori)-Rλ(co)-Rzλ(co) <br> | + | <b>λLysis_Sm(o_c_c)</b>: <i>Sλmut(ori)-Rλ(co)-Rzλ(co)</i> <br> |
− | <b>λLysis_Sm(c_c_c)</b>: Sλmut(co)-Rλ(co)-Rzλ(co) <br> | + | <b>λLysis_Sm(c_c_c)</b>: <i>Sλmut(co)-Rλ(co)-Rzλ(co)</i> <br> |
[x/y]: restriction sites digested: E, EcoRI; X, XbaI; S, SpeI; P, PstI <br> | [x/y]: restriction sites digested: E, EcoRI; X, XbaI; S, SpeI; P, PstI <br> | ||
Line 414: | Line 414: | ||
<td>Wednesday 10</td> | <td>Wednesday 10</td> | ||
<td></td> | <td></td> | ||
− | <td><li>Extraction of the plasmid pGOv4-Rzλ(co)</li></td> | + | <td><li>Extraction of the plasmid pGOv4-<i>Rzλ(co)</i></li></td> |
− | <td><li>Extraction of the plasmid pGOv4-Rλ(co)</li></td> | + | <td><li>Extraction of the plasmid pGOv4-<i>Rλ(co)</i></li></td> |
<td></td> | <td></td> | ||
</tr> | </tr> | ||
Line 421: | Line 421: | ||
<td>Thursday 11</td> | <td>Thursday 11</td> | ||
<td><li>Preparation of the plasmid pSB1C3</li> <li>Restriction digestion with [E/P]</li></td> | <td><li>Preparation of the plasmid pSB1C3</li> <li>Restriction digestion with [E/P]</li></td> | ||
− | <td><li>Restriction digestion with [E/P] on the plasmid pGOv4-Rzλ(co) </li> <li>Gel purification</li></td> | + | <td><li>Restriction digestion with [E/P] on the plasmid pGOv4-<i>Rzλ(co)</i> </li> <li>Gel purification</li></td> |
− | <td><li>Restriction digestion with [E/P] on the plasmid pGOv4-Rλ(co) </li> <li>Gel purification</li></td> | + | <td><li>Restriction digestion with [E/P] on the plasmid pGOv4-<i>Rλ(co)</i> </li> <li>Gel purification</li></td> |
<td></td> | <td></td> | ||
</tr> | </tr> | ||
Line 428: | Line 428: | ||
<td>Friday 12</td> | <td>Friday 12</td> | ||
<td></td> | <td></td> | ||
− | <td><li>Ligation of the [E/P] digested Rzλ(co) insert (from June 11) with the [E/P] digested vector pSB1C3</li> <li>Transformation of the ligaiton product into competent <i>E. coli</i> cells</li></td> | + | <td><li>Ligation of the [E/P] digested <i>Rzλ(co)</i> insert (from June 11) with the [E/P] digested vector pSB1C3</li> <li>Transformation of the ligaiton product into competent <i>E. coli</i> cells</li></td> |
− | <td></td> | + | <td><li>Redo 11/6’s work</li></td> |
<td></td> | <td></td> | ||
</tr> | </tr> | ||
Line 453: | Line 453: | ||
<td></td> | <td></td> | ||
<td><li>Colony PCR on June 12 transformed cells (Result: no bands amplified)</li></td> | <td><li>Colony PCR on June 12 transformed cells (Result: no bands amplified)</li></td> | ||
− | <td><li>Ligation of the [E/P] digested Rλ(co) insert (from June 12) into the [E/P] digested vector pSB1C3</li> <li>Transformation of the ligation product into competent <i>E. coli</i> cells</li></td> | + | <td><li>Ligation of the [E/P] digested <i>Rλ(co)</i> insert (from June 12) into the [E/P] digested vector pSB1C3</li> <li>Transformation of the ligation product into competent <i>E. coli</i> cells</li></td> |
<td></td> | <td></td> | ||
</tr> | </tr> | ||
Line 467: | Line 467: | ||
<td>Thursday 18</td> | <td>Thursday 18</td> | ||
<td></td> | <td></td> | ||
− | <td><li>Redo the ligation of the [E/P] digested Rzλ(co) insert into the [E/P] digested vector pSB1C3 </li> <li>Transformation of the ligation product into competent <i>E. coli</i> cells</li></td> | + | <td><li>Redo the ligation of the [E/P] digested <i>Rzλ(co)</i> insert into the [E/P] digested vector pSB1C3 </li> <li>Transformation of the ligation product into competent <i>E. coli</i> cells</li></td> |
− | <td><li>Redo the extraction of the plasmid pGOv4-Rλ(co)</li> <li>Restriction digestion with [E/P]</li></td> | + | <td><li>Redo the extraction of the plasmid pGOv4-<i>Rλ(co)</i></li> <li>Restriction digestion with [E/P]</li></td> |
<td></td> | <td></td> | ||
</tr> | </tr> | ||
Line 502: | Line 502: | ||
<td></td> | <td></td> | ||
<td></td> | <td></td> | ||
− | <td><li>Redo the extraction of the plasmid pGOv4-Rλ(co)</li> <li>Restriction digestion with [E/P]</li> <li>Gel purification</li></td> | + | <td><li>Redo the extraction of the plasmid pGOv4-<i>Rλ(co)</i></li> <li>Restriction digestion with [E/P]</li> <li>Gel purification</li></td> |
<td></td> | <td></td> | ||
</tr> | </tr> | ||
Line 517: | Line 517: | ||
<td></td> | <td></td> | ||
<td></td> | <td></td> | ||
− | <td><li>Ligation of the [E/P] digested Rλ(co) insert into the [E/P] digested pSB1C3 vector</li> <li>Transformation of the ligation product into competent <i>E. coli</i> cells</li></td> | + | <td><li>Ligation of the [E/P] digested <i>Rλ(co)</i> insert into the [E/P] digested pSB1C3 vector</li> <li>Transformation of the ligation product into competent <i>E. coli</i> cells</li></td> |
<td></td> | <td></td> | ||
</tr> | </tr> | ||
Line 543: | Line 543: | ||
<td>Monday 29</td> | <td>Monday 29</td> | ||
<td></td> | <td></td> | ||
− | <td><li>Extraction of the plasmid pSB1C3-Rzλ(co) (from June 18 transformed cells)</li></td> | + | <td><li>Extraction of the plasmid pSB1C3-<i>Rzλ(co)</i> (from June 18 transformed cells)</li></td> |
<td><li>Colony PCR on June 25 transformed cells (Result: again showed no bands)</li></td> | <td><li>Colony PCR on June 25 transformed cells (Result: again showed no bands)</li></td> | ||
<td></td> | <td></td> | ||
Line 551: | Line 551: | ||
<td></td> | <td></td> | ||
<td></td> | <td></td> | ||
− | <td><li>Sequencing result confirmed the presence of Rλ(co) insert in pSB1C3</li></td> | + | <td><li>Sequencing result confirmed the presence of <i>Rλ(co)</i> insert in pSB1C3</li></td> |
<td></td> | <td></td> | ||
</tr> | </tr> | ||
Line 562: | Line 562: | ||
<tr> | <tr> | ||
<td>Thursday 2</td> | <td>Thursday 2</td> | ||
− | <td><li>PCR amplification of the BBa_K124017 SλWT(ori) gene</li></td> | + | <td><li>PCR amplification of the BBa_K124017 <i>SλWT(ori)</i> gene</li></td> |
− | <td><li><font color="#D633FF">PCR amplification of the BBa_K124017 Rλ(ori) gene</font></li> <li><font color="#D633FF">Rλ(ori) PCR product purification</font></li> <li><font color="#0099FF">Transformation of pGOv4-SλWT(co) into competent <i>E. coli</i> cells</font></li></td> | + | <td><li><font color="#D633FF">PCR amplification of the BBa_K124017 <i>Rλ(ori)</i> gene</font></li> <li><font color="#D633FF"><i>Rλ(ori)</i> PCR product purification</font></li> <li><font color="#0099FF">Transformation of pGOv4-<i>SλWT(co)</i> into competent <i>E. coli</i> cells</font></li></td> |
− | <td><li><font color="#D633FF">PCR amplification of the BBa_K124017 Rzλ(ori) gene</font></li> <li><font color="#D633FF">Rzλ(ori) PCR product purification</font></li> <li><font color="#D633FF">Restriction digestion with [E/P]</font></li></td> | + | <td><li><font color="#D633FF">PCR amplification of the BBa_K124017 <i>Rzλ(ori)</i> gene</font></li> <li><font color="#D633FF"><i>Rzλ(ori)</i> PCR product purification</font></li> <li><font color="#D633FF">Restriction digestion with [E/P]</font></li></td> |
− | <td><li>PCR amplification of the BBa_K124017 Rλ(ori) gene</li> <li>Rλ(ori) PCR product purification</li></td> | + | <td><li>PCR amplification of the BBa_K124017 <i>Rλ(ori)</i> gene</li> <li><i>Rλ(ori)</i> PCR product purification</li></td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>Friday 3</td> | <td>Friday 3</td> | ||
<td><li>Restriction digestion of pSB1C3 vector with [E/X]</li></td> | <td><li>Restriction digestion of pSB1C3 vector with [E/X]</li></td> | ||
− | <td><li>Restriction digestion of Rλ(co) with [E/P]</li> <li>Gel purification</li></td> | + | <td><li>Restriction digestion of <i>Rλ(co)</i> with [E/P]</li> <li>Gel purification</li></td> |
− | <td>[1]<li><font color="#D633FF">Gel purification of July 2 [E/P] digested Rzλ(ori)</font></li> <li>Ligation of the [E/P] digested <font color="#D633FF">Rzλ(ori) insert </font>into [E/P] digested pSB1C3 vector</li> [2] <li><font color="#0099FF">Extraction of the plasmid pSB1C3-Rλ(co) (from June 25 transformed cells)</font></li> <li><font color="#0099FF">Restriction digestion with [S/P]</li> <li>Gel purification</font></li> [3] <li><font color="#D633FF">Restriction digestion of June 2 Rzλ(ori) with [X/P] & Heat kill</font></li> <li><font color="#00CC00">Ligation of [X/P] digested Rzλ(ori) insert into [S/P] digested pSB1C3-Rλ(co) vector</li></td> | + | <td>[1]<li><font color="#D633FF">Gel purification of July 2 [E/P] digested <i>Rzλ(ori)</i></font></li> <li>Ligation of the [E/P] digested <font color="#D633FF"><i>Rzλ(ori)</i> insert </font>into [E/P] digested pSB1C3 vector</li> [2] <li><font color="#0099FF">Extraction of the plasmid pSB1C3-<i>Rλ(co)</i> (from June 25 transformed cells)</font></li> <li><font color="#0099FF">Restriction digestion with [S/P]</li> <li>Gel purification</font></li> [3] <li><font color="#D633FF">Restriction digestion of June 2 <i>Rzλ(ori)</i> with [X/P] & Heat kill</font></li> <li><font color="#00CC00">Ligation of [X/P] digested Rzλ(ori) insert into [S/P] digested pSB1C3-<i>Rλ(co)</i> vector</li></td> |
− | <td><li>Extraction of the plasmid pSB1C3-Rλ(co) (from Gp3 June 25 transformed cells)</li></td> | + | <td><li>Extraction of the plasmid pSB1C3-<i>Rλ(co)</i> (from Gp3 June 25 transformed cells)</li></td> |
</tr> | </tr> | ||
</table> | </table> | ||
Line 589: | Line 589: | ||
<tr> | <tr> | ||
<td>Monday 6</td> | <td>Monday 6</td> | ||
− | <td><li>Gel purification of the [X/P] digested pSB1C3 vector isolated on July 3</li> <li><font color="#D633FF">PCR amplification of the BBa_K124017 Rzλ(ori) gene using a F-primer with SacII restriction site</font></li></td> | + | <td><li>Gel purification of the [X/P] digested pSB1C3 vector isolated on July 3</li> <li><font color="#D633FF">PCR amplification of the BBa_K124017 <i>Rzλ(ori)</i> gene using a F-primer with SacII restriction site</font></li></td> |
<td></td> | <td></td> | ||
− | <td><li><font color="#00CC00">Transformation of the ligation product pSB1C3-Rλ(co)-Rzλ(ori) (from July 3 [3])</font></li></td> | + | <td><li><font color="#00CC00">Transformation of the ligation product pSB1C3-<i>Rλ(co)-Rzλ(ori)</i> (from July 3 [3])</font></li></td> |
− | <td><li>Restriction digestion of July 3 Rλ(co) with [S/P]</li><li>Gel purification</li> <br> <li>Restriction digestion of Rzλ(co) (from Gp2 June 29) with [X/P]</li><li>Gel purification</li></td> | + | <td><li>Restriction digestion of July 3 <i>Rλ(co)</i> with [S/P]</li><li>Gel purification</li> <br> <li>Restriction digestion of <i>Rzλ(co)</i> (from Gp2 June 29) with [X/P]</li><li>Gel purification</li></td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>Tuesday 7</td> | <td>Tuesday 7</td> | ||
− | <td><li><font color="#D633FF">Gel electrophoresis of the SacII digested Rzλ(ori) PCR amplicon</font></li> <li><font color="#0099FF">PCR amplification of SλWT(ori) using a new primer pair</font></li></td> | + | <td><li><font color="#D633FF">Gel electrophoresis of the SacII digested <i>Rzλ(ori)</i> PCR amplicon</font></li> <li><font color="#0099FF">PCR amplification of <i>SλWT(ori)</i> using a new primer pair</font></li></td> |
− | <td><li><font color="#0099FF">Extraction of the plasmid pGOv4-SλWT(co)</font></li><li><font color="#D633FF">Restriction digestion with [X] on July 3 [P] digested Rλ(ori)</font></li></td> | + | <td><li><font color="#0099FF">Extraction of the plasmid pGOv4-<i>SλWT(co)</i></font></li><li><font color="#D633FF">Restriction digestion with [X] on July 3 [P] digested <i>Rλ(ori)</i></font></li></td> |
− | <td><li><font color="#00CC00">Colony PCR on July 6 Rλ(co)-Rzλ(ori) clones showed no bands</li> <li>Redo July 3’s ligation of [X/P] digested Rzλ(ori) insert into [S/P] digested pSB1C3-Rλ(co) vector</li> <li>Transformation</font></li></td> | + | <td><li><font color="#00CC00">Colony PCR on July 6 <i>Rλ(co)-Rzλ(ori)</i> clones showed no bands</li> <li>Redo July 3’s ligation of [X/P] digested <i>Rzλ(ori)</i> insert into [S/P] digested pSB1C3-<i>Rλ(co)</i> vector</li> <li>Transformation</font></li></td> |
− | <td><li>Ligation of the [X/P] digested Rzλ(co) insert into the [S/P] digested pSB1C3-Rλ(co) vector</li> <li>Transformation</li></td> | + | <td><li>Ligation of the [X/P] digested <i>Rzλ(co)</i> insert into the [S/P] digested pSB1C3-<i>Rλ(co)</i> vector</li> <li>Transformation</li></td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>Wednesday 8</td> | <td>Wednesday 8</td> | ||
− | <td><li><font color="#D633FF">PCR purification of July 6 SacII Rzλ(ori) PCR product</font></li></td> | + | <td><li><font color="#D633FF">PCR purification of July 6 SacII <i>Rzλ(ori)</i> PCR product</font></li></td> |
− | <td><li><font color="#D633FF">Ligation of [X/P] digested Rλ(ori) into Gp1 July 6 [X/P]</li><li>Transformation of 7/7’s ligation product Rλ(ori) in pSB1C3</font></li></td> | + | <td><li><font color="#D633FF">Ligation of [X/P] digested <i>Rλ(ori)</i> into Gp1 July 6 [X/P]</li><li>Transformation of 7/7’s ligation product <i>Rλ(ori)</i> in pSB1C3</font></li></td> |
− | <td><li><font color="#00CC00">Colony PCR on July 7 Rλ(co)-Rzλ(ori) clones still showed no bands</font></li></td> | + | <td><li><font color="#00CC00">Colony PCR on July 7 <i>Rλ(co)-Rzλ(ori)</i> clones still showed no bands</font></li></td> |
− | <td><li>Colony PCR on July 7 Rλ(co)-Rzλ(co) clones showed no bands</li></td> | + | <td><li>Colony PCR on July 7 <i>Rλ(co)-Rzλ(co)</i> clones showed no bands</li></td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>Thursday 9</td> | <td>Thursday 9</td> | ||
− | <td><li><font color="#D633FF">Restriction digestion of SacII Rzλ(ori) with [SacII/P]</font></li><li><font color="#00CC00">Restriction digestion of Gp2 July 7 SλWT(co) with [E/S]</font></li><li><font color="#0099FF">Gel electrophoresis of July 7 SλWT(ori)</li><li>PCR purification of SλWT(ori)</font></li></td> | + | <td><li><font color="#D633FF">Restriction digestion of SacII <i>Rzλ(ori)</i> with [SacII/P]</font></li><li><font color="#00CC00">Restriction digestion of Gp2 July 7 <i>SλWT(co)</i> with [E/S]</font></li><li><font color="#0099FF">Gel electrophoresis of July 7 <i>SλWT(ori)</i></li><li>PCR purification of <i>SλWT(ori)</i></font></li></td> |
− | <td><li><font color="#D633FF">Colony PCR on Rλ(ori) (from July 8 transformed cells)</font></li> <li><font color="#0099FF">Restriction digestion of July 7 SλWT(co) with [E/P]</font></li></td> | + | <td><li><font color="#D633FF">Colony PCR on <i>Rλ(ori)</i> (from July 8 transformed cells)</font></li> <li><font color="#0099FF">Restriction digestion of July 7 <i>SλWT(co)</i> with [E/P]</font></li></td> |
− | <td><li><font color="#D633FF">Ligation of [X/P] July 3 digested Rzλ(ori) insert into [X/P] digested pSB1C3 vector</li> <li>Transformation</font></li></td> | + | <td><li><font color="#D633FF">Ligation of [X/P] July 3 digested <i>Rzλ(ori)</i> insert into [X/P] digested pSB1C3 vector</li> <li>Transformation</font></li></td> |
<td></td> | <td></td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>Friday 10</td> | <td>Friday 10</td> | ||
− | <td><li>Restriction digestion of SλWT(ori) with [X/P] & Gel purification</li><li>Ligation of [X/P] digested SλWT(ori) insert into July 6 [X/P] digested pSB1C3 vector</li><li>Transformation</li><li>Gel purification of July 9 [E/S] digested SλWT(co)</li></td> | + | <td><li>Restriction digestion of <i>SλWT(ori)</i> with [X/P] & Gel purification</li><li>Ligation of [X/P] digested <i>SλWT(ori)</i> insert into July 6 [X/P] digested pSB1C3 vector</li><li>Transformation</li><li>Gel purification of July 9 [E/S] digested <i>SλWT(co)</i></li></td> |
− | <td><li><font color="#0099FF">Gel purification of July 9 [E/P] digested SλWT(co)</font></li></td> | + | <td><li><font color="#0099FF">Gel purification of July 9 [E/P] digested <i>SλWT(co)</i></font></li></td> |
− | <td><li><font color="#D633FF">Colony PCR on July 9 Rzλ(ori) clones showed no bands</font></li><li><font color="#00CC00">Restriction analysis on July 7 transformed cells</li><li>Another redo of the ligation of July 3 [X/P] digested Rzλ(ori) insert into July 3 [S/P] digested pSB1C3-Rλ(co)</li><li>Transformation</font></li></td> | + | <td><li><font color="#D633FF">Colony PCR on July 9 <i>Rzλ(ori)</i> clones showed no bands</font></li><li><font color="#00CC00">Restriction analysis on July 7 transformed cells</li><li>Another redo of the ligation of July 3 [X/P] digested <i>Rzλ(ori)</i> insert into July 3 [S/P] digested pSB1C3-<i>Rλ(co)</i></li><li>Transformation</font></li></td> |
− | <td><li>Extraction of the plasmid pSB1C3-Rλ(co)-Rzλ(co) (from July 7 transformed cells)</li> <li>Restriction analysis of Rλ(co)-Rzλ(co) showed bands of the expected size</li></td> | + | <td><li>Extraction of the plasmid pSB1C3-<i>Rλ(co)-Rzλ(co)</i> (from July 7 transformed cells)</li> <li>Restriction analysis of <i>Rλ(co)-Rzλ(co)</i> showed bands of the expected size</li></td> |
</tr> | </tr> | ||
</table> | </table> | ||
Line 639: | Line 639: | ||
<tr> | <tr> | ||
<td>Monday 13</td> | <td>Monday 13</td> | ||
− | <td><li><font color="#0099FF">Colony PCR on SλWT(ori) clones showed bands of the expected size</font></li><li><font color="#00CC00">Ligation of [E/S] digested SλWT(co) insert into pSB1C3 vector</li><li>Transformation</font></li></td> | + | <td><li><font color="#0099FF">Colony PCR on <i>SλWT(ori)</i> clones showed bands of the expected size</font></li><li><font color="#00CC00">Ligation of [E/S] digested <i>SλWT(co)</i> insert into pSB1C3 vector</li><li>Transformation</font></li></td> |
− | <td><li><font color="#D633FF">Extraction of the plasmid pSB1C3-Rλ(ori)</li><li>Restriction digestion with [X/P]</li><li>Gel purification </font></li></td> | + | <td><li><font color="#D633FF">Extraction of the plasmid pSB1C3-<i>Rλ(ori)</i></li><li>Restriction digestion with [X/P]</li><li>Gel purification </font></li></td> |
− | <td><li><font color="#00CC00">Colony PCR on 10/7 transformed cells pSB1C3-Rλ(co)-Rzλ(ori)</font></li></td> | + | <td><li><font color="#00CC00">Colony PCR on 10/7 transformed cells pSB1C3-<i>Rλ(co)-Rzλ(ori)</i></font></li></td> |
<td></td> | <td></td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>Tuesday 14</td> | <td>Tuesday 14</td> | ||
− | <td><li><font color="#00CC00">Colony PCR on SλWT(co) clones showed bands of the expected size</font></li> <li><font color="#FF3333"> Restriction digestion of Gp2 Rλ(ori) with [X/SacII]</font></li></td> | + | <td><li><font color="#00CC00">Colony PCR on <i>SλWT(co)</i> clones showed bands of the expected size</font></li> <li><font color="#FF3333"> Restriction digestion of Gp2 <i>Rλ(ori)</i> with [X/SacII]</font></li></td> |
− | <td><li><font color="#D633FF">Restriction analysis of [X/P] digested Rλ(ori)</font></li><li>Restriction digestion of Rzλ(co) with [X/P]</li><li>Gel purification</li></td> | + | <td><li><font color="#D633FF">Restriction analysis of [X/P] digested <i>Rλ(ori)</i></font></li><li>Restriction digestion of <i>Rzλ(co)</i> with [X/P]</li><li>Gel purification</li></td> |
<td></td> | <td></td> | ||
− | <td><li>Extraction of the plasmid pSB1C3-Rλ(co)-Rzλ(co)</li> <li>Restriction digestion with [X/P]</li></td> | + | <td><li>Extraction of the plasmid pSB1C3-<i>Rλ(co)-Rzλ(co)</i></li> <li>Restriction digestion with [X/P]</li></td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>Wednesday 15</td> | <td>Wednesday 15</td> | ||
− | <td><li><font color="#FF3333">Gel purification of July 14 [X/SacII] digested Rλ(ori)</font></li><br><li><font color="#FF9900">Ligation between</font> <font color="#00CC00">[E/S] digested SλWT(co)</font>, <font color="#FF3333">[X/SacII] digested Rλ(ori)</font>, <font color="#D633FF">[SacII/P] digested Rzλ(ori)</font> and <font color="#FF9900">[E/P] digested pSB1C3 backbone</li><li>Transformation</font></li></td> | + | <td><li><font color="#FF3333">Gel purification of July 14 [X/SacII] digested <i>Rλ(ori)</i></font></li><br><li><font color="#FF9900">Ligation between</font> <font color="#00CC00">[E/S] digested <i>SλWT(co)</i></font>, <font color="#FF3333">[X/SacII] digested <i>Rλ(ori)</i></font>, <font color="#D633FF">[SacII/P] digested <i>Rzλ(ori)</i></font> and <font color="#FF9900">[E/P] digested pSB1C3 backbone</li><li>Transformation</font></li></td> |
− | <td><li><font color="#D633FF">Extraction of the plasmid pSB1C3-Rλ(ori)</li><li>Restriction digestion with [S/P]</font></li></td> | + | <td><li><font color="#D633FF">Extraction of the plasmid pSB1C3-<i>Rλ(ori)</i></li><li>Restriction digestion with [S/P]</font></li></td> |
− | <td><li>Extraction of the plasmid pSB1C3-Rλ(co)</li><li>Restriction digestion of Rλ(co) with [E/S]</li><li>Restriction digestion of [E/X] digested Rzλ(ori) with CIP & Gel purification</li><li>Ligation of the [E/S] digested Rλ(co) insert into [E/X] digested pSB1C3-Rzλ(ori)</li><li>Transformation</li></td> | + | <td><li>Extraction of the plasmid pSB1C3-<i>Rλ(co)</i></li><li>Restriction digestion of <i>Rλ(co)</i> with [E/S]</li><li>Restriction digestion of [E/X] digested <i>Rzλ(ori)</i> with CIP & Gel purification</li><li>Ligation of the [E/S] digested <i>Rλ(co)</i> insert into [E/X] digested pSB1C3-<i>Rzλ(ori)</i></li><li>Transformation</li></td> |
− | <td><li>Gel purification of July 14 [X/P] digested Rλ(co)-Rzλ(co)</li></td> | + | <td><li>Gel purification of July 14 [X/P] digested <i>Rλ(co)-Rzλ(co)</i></li></td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>Thursday 16</td> | <td>Thursday 16</td> | ||
− | <td><li><font color="#FF9900">Colony PCR on SλWT(co)- Rλ(ori)-Rzλ(ori) showed bands believed to be the desired insert</font></li></td> | + | <td><li><font color="#FF9900">Colony PCR on <i>SλWT(co)- Rλ(ori)-Rzλ(ori)</i> showed bands believed to be the desired insert</font></li></td> |
− | <td><li><font color="#D633FF">Gel purification of July 15 [S/P] digested Rλ(ori)</font></li> <li>Ligation of the [X/P] digested Rzλ(co) insert into <font color="#D633FF">[S/P] digestd pSB1C3-Rλ(ori) vector</font></li></td> | + | <td><li><font color="#D633FF">Gel purification of July 15 [S/P] digested <i>Rλ(ori)</i></font></li> <li>Ligation of the [X/P] digested <i>Rzλ(co)</i> insert into <font color="#D633FF">[S/P] digestd pSB1C3-<i>Rλ(ori)</i> vector</font></li></td> |
− | <td><li>Colony PCR on July 15 transformed cells</li> <li>Redo the ligation of [E/S] digested Rλ(co) insert into [E/X] digested pSB1C3-Rzλ(ori)</li> <li>Transformation</li></td> | + | <td><li>Colony PCR on July 15 transformed cells</li> <li>Redo the ligation of [E/S] digested <i>Rλ(co)</i> insert into [E/X] digested pSB1C3-<i>Rzλ(ori)</i></li> <li>Transformation</li></td> |
<td></td> | <td></td> | ||
</tr> | </tr> | ||
Line 670: | Line 670: | ||
<td>Friday 17</td> | <td>Friday 17</td> | ||
<td></td> | <td></td> | ||
− | <td><li>Transformation of July 16 ligation product pSB1C3-Rλ(ori)-Rzλ(co)</li></td> | + | <td><li>Transformation of July 16 ligation product pSB1C3-<i>Rλ(ori)-Rzλ(co)</i></li></td> |
− | <td><li>Colony PCR on 16/7 transformed cells</li><li>Restriction digestion of Sλ(co) with [E/S] & Gel purification</li><li>Redo the ligation of [E/S] digested Rλ(co) insert into [E/X] digested pSB1C3-Rzλ(ori)</li><li>Transformation</li></td> | + | <td><li>Colony PCR on 16/7 transformed cells</li><li>Restriction digestion of <i>Sλ(co)</i> with [E/S] & Gel purification</li><li>Redo the ligation of [E/S] digested <i>Rλ(co)</i> insert into [E/X] digested pSB1C3-<i>Rzλ(ori)</i></li><li>Transformation</li></td> |
<td></td> | <td></td> | ||
</tr> | </tr> | ||
Line 680: | Line 680: | ||
<td></td> | <td></td> | ||
<td></td> | <td></td> | ||
− | <td><li><font color="#D633FF">Extraction of the plasmids pSB1C3-Sλmut(ori) and pSB1C3-Sλmut(co)</li><li>Restriction digestion with [S/P]</font></li></td> | + | <td><li><font color="#D633FF">Extraction of the plasmids pSB1C3-<i>Sλmut(ori)</i> and pSB1C3-<i>Sλmut(co)</i></li><li>Restriction digestion with [S/P]</font></li></td> |
</table> | </table> | ||
<br> | <br> | ||
− | <font size="3">DNA sequencing results later revealed that the genes amplified from the BioBrick, BBa_K124017, did not contain the desired sequences. Genomic DNA from E. coli BL21(DE3) will be used as template (starting from week 10) to create new λ(ori) related BioBricks. Gene sequences of the λ(co) clones were correct.</font> | + | <font size="3">DNA sequencing results later revealed that the genes amplified from the BioBrick, BBa_K124017, did not contain the desired sequences. Genomic DNA from <i>E. coli</i> BL21(DE3) will be used as template (starting from week 10) to create new λ(ori) related BioBricks. Gene sequences of the λ(co) clones were correct.</font> |
<br><br> | <br><br> | ||
Line 703: | Line 703: | ||
<td></td> | <td></td> | ||
<td></td> | <td></td> | ||
− | <td><li><font color="#D633FF"> Gel purification of July 18 [S/P] digested pSB1C3-Sλmut(ori) and pSB1C3-Sλmut(co)</font></li><li><font color="#0099FF">Ligation of July 15 [X/P] digested Rλ(co)-Rzλ(co) insert into the [S/P] digested pSB1C3-Sλmut(ori) vector</font></li><li><font color="#FF9900"> Ligation of July 15 [X/P] digested Rλ(co)-Rzλ(co) insert into the [S/P] digested pSB1C3-Sλmut(co) vector</font></li></td> | + | <td><li><font color="#D633FF"> Gel purification of July 18 [S/P] digested pSB1C3-<i>Sλmut(ori)</i> and pSB1C3-<i>Sλmut(co)</i></font></li><li><font color="#0099FF">Ligation of July 15 [X/P] digested <i>Rλ(co)-Rzλ(co)</i> insert into the [S/P] digested pSB1C3-<i>Sλmut(ori)</i> vector</font></li><li><font color="#FF9900"> Ligation of July 15 [X/P] digested <i>Rλ(co)-Rzλ(co)</i> insert into the [S/P] digested pSB1C3-<i>Sλmut(co)</i> vector</font></li></td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>Tuesday 21</td> | <td>Tuesday 21</td> | ||
<td></td> | <td></td> | ||
− | <td><li>Transformation of pGOv4- | + | <td><li>Transformation of pGOv4-Ribo_<i>Sλ(co)</i>, pGOv4-Ribo_<i>Rλ(co)</i> and pGOv4-Ribo_<i>Rzλ(co)</i> into competent <i>E. coli</i> cells</li><li>Transformation of pGOv4-<i>SλWT(co)-Rλ(co)-Rzλ(co)</i> (λLysis(c_c_c))</li></td> |
<td></td> | <td></td> | ||
− | <td><li><font color="#0099FF">Transformation of the pSB1C3-Sλmut(ori)-Rλ(co)-Rzλ(co) (λLysis_Sm(o_c_c))</font></li> <li><font color="#FF9900"> Transformation of the pSB1C3-Sλmut(co)-Rλ(co)-Rzλ(co) (λLysis_Sm(c_c_c))</font></li></td> | + | <td><li><font color="#0099FF">Transformation of the pSB1C3-<i>Sλmut(ori)-Rλ(co)-Rzλ(co)</i> (λLysis_Sm(o_c_c))</font></li> <li><font color="#FF9900"> Transformation of the pSB1C3-<i>Sλmut(co)-Rλ(co)-Rzλ(co)</i> (λLysis_Sm(c_c_c))</font></li></td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>Wednesday 22</td> | <td>Wednesday 22</td> | ||
− | <td><li><font color=”#3333FF”>Genomic DNA extraction of BL21(DE3) <i>E. coli</i> cells</li><li>PCR amplification of the Sλ(ori), Rλ(ori) and Rzλ(ori) genes using the extracted genomic DNA as template</font></li></td> | + | <td><li><font color=”#3333FF”>Genomic DNA extraction of BL21(DE3) <i>E. coli</i> cells</li><li>PCR amplification of the <i>Sλ(ori)</i>, <i>Rλ(ori)</i> and <i>Rzλ(ori)</i> genes using the extracted genomic DNA as template</font></li></td> |
<td></td> | <td></td> | ||
− | <td><li><font color="#FF00FF">Redo Gp1 July 21 transformation of the pGOv4- | + | <td><li><font color="#FF00FF">Redo Gp1 July 21 transformation of the pGOv4-Ribo_<i>Sλ(co)</i> and pGOv4-<i>SλWT(co)-Rλ(co)-Rzλ(co)</i> (λLysis (c_c_c)) into competent <i>E. coli</i> cells</font></li></td> |
− | <td><li><font color="#0099FF"> Colony PCR on λLysis_Sm(o_c_c) clones showed bands of the expected size</font></li> <br> <li><font color="#FF9900">Colony PCR on c clones revealed that that the colonies contained inserts of the wrong sizes</li><li>Redo the restriction digestion of Rλ(co)-Rzλ(co) with [X/P] & Gel purification</font></li></td> | + | <td><li><font color="#0099FF"> Colony PCR on λLysis_Sm(o_c_c) clones showed bands of the expected size</font></li> <br> <li><font color="#FF9900">Colony PCR on c clones revealed that that the colonies contained inserts of the wrong sizes</li><li>Redo the restriction digestion of <i>Rλ(co)-Rzλ(co)</i> with [X/P] & Gel purification</font></li></td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>Thursday 23</td> | <td>Thursday 23</td> | ||
− | <td><li><font color="#FF00FF">Extraction of the plasmids pGOv4- | + | <td><li><font color="#FF00FF">Extraction of the plasmids pGOv4-Ribo_<i>Sλ(co)</i>, pGOv4-Ribo_<i>Rλ(co)</i> and pGOv4-Ribo_<i>Rzλ(co)</i></li><li>Restriction digestion of the extracted plasmids with [E/P]</li><li>Gel purification</font></li></td> |
<td></td> | <td></td> | ||
<td><li><font color="#FF00FF">Extraction of the plasmid pGOv4-λLysis(c_c_c)</li> <li>Restriction digestion with [E/P]</font></li></td> | <td><li><font color="#FF00FF">Extraction of the plasmid pGOv4-λLysis(c_c_c)</li> <li>Restriction digestion with [E/P]</font></li></td> | ||
− | <td><li><font color="#FF9900">Redo the ligation of the [X/P] digested Rλ(co)-Rzλ(co) insert into the [S/P] digested pSB1C3-Sλmut(co) vector</li><li>Transformation</font></li></td> | + | <td><li><font color="#FF9900">Redo the ligation of the [X/P] digested <i>Rλ(co)-Rzλ(co)</i> insert into the [S/P] digested pSB1C3-<i>Sλmut(co)</i> vector</li><li>Transformation</font></li></td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>Friday 24</td> | <td>Friday 24</td> | ||
− | <td><li><font color="#FF00FF">Ligation of the [E/P] digested | + | <td><li><font color="#FF00FF">Ligation of the [E/P] digested Ribo_<i>Sλ(co)</i> and Ribo_<i>Sλ(co)</i> inserts into [E/P] digested pSB1C3 respectively</li><li>Transformation</font></li><br><li><font color="#3333FF">Restriction digestion of the <i>Sλ(ori)</i>, <i>Rλ(ori)</i> and <i>Rzλ(ori)</i> amplicons with [X/P]</font></li></td> |
<td></td> | <td></td> | ||
<td></td> | <td></td> | ||
− | <td><li><font color="#FF9900">Colony PCR on the new λLysis_Sm(c_c_c) clones showed bands of the expected size. However, later sequencing result revealed that the colony was actually a mixed one with both the correct plasmid and the self-ligated RRz(co) plasmid</font></li></td> | + | <td><li><font color="#FF9900">Colony PCR on the new λLysis_Sm(c_c_c) clones showed bands of the expected size. However, later sequencing result revealed that the colony was actually a mixed one with both the correct plasmid and the self-ligated <i>RRz(co)</i> plasmid</font></li></td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>Saturday 25</td> | <td>Saturday 25</td> | ||
− | <td><li><font color="#FF00FF">Colony PCR on July 24 transformed | + | <td><li><font color="#FF00FF">Colony PCR on July 24 transformed Ribo_<i>Sλ</i> and Ribo_<i>Rλ</i> clones</font></li><br><li><font color="#3333FF">Ligation of the [X/P] digested <i>Sλ(ori)</i>, <i>Rλ(ori)</i> and <i>Rzλ(ori)</i> inserts into [X/P] digested pSB1C3 respectively</li><li>Transformation</font></li></td> |
<td></td> | <td></td> | ||
<td></td> | <td></td> | ||
Line 760: | Line 760: | ||
<tr> | <tr> | ||
<td>Tuesday 28</td> | <td>Tuesday 28</td> | ||
− | <td><li><font color="#3333FF">Colony PCR on Sλ(ori), Rλ(ori) and Rzλ(ori) clones showed bands of the expected size</font></li></td> | + | <td><li><font color="#3333FF">Colony PCR on <i>Sλ(ori)</i>, <i>Rλ(ori</i>) and <i>Rzλ(ori)</i> clones showed bands of the expected size</font></li></td> |
<td></td> | <td></td> | ||
− | <td><li>Restriction digestion of | + | <td><li>Restriction digestion of Ribo_<i>Rλ(co)</i> with [E/P]</li></td> |
<td></td> | <td></td> | ||
</tr> | </tr> | ||
Line 785: | Line 785: | ||
<td></td> | <td></td> | ||
<td></td> | <td></td> | ||
− | <td>Redo the restriction digestion of | + | <td>Redo the restriction digestion of Ribo_<i>Rλ(co)</i> with [E/P]</td> |
<td><li><font color="#0099FF"> Redo the extraction of the plasmid pSB1C3-λLysis_Sm(o_c_c)</li><li>Redo the restriction digestion with [X/P]</li><li>Gel purification</font></li></td> | <td><li><font color="#0099FF"> Redo the extraction of the plasmid pSB1C3-λLysis_Sm(o_c_c)</li><li>Redo the restriction digestion with [X/P]</li><li>Gel purification</font></li></td> | ||
</tr> | </tr> | ||
Line 812: | Line 812: | ||
<td></td> | <td></td> | ||
<td></td> | <td></td> | ||
− | <td><li><font color="#FF9900">Redo the ligation of the [X/P] digested Rλ(co)-Rzλ(co) insert into the [S/P] digested pSB1C3-Sλmut(co) vector</font></li> | + | <td><li><font color="#FF9900">Redo the ligation of the [X/P] digested <i>Rλ(co)-Rzλ(co)</i> insert into the [S/P] digested pSB1C3-<i>Sλmut(co)</i> vector</font></li> |
<br> | <br> | ||
− | <li><font color="#FF3333"> Extraction of the plasmid pGOv4-lacIq (pLacIQ_LacI_L8-UV5 lac)</font></li> | + | <li><font color="#FF3333"> Extraction of the plasmid pGOv4-<i>lacIq</i> (pLacIQ_LacI_L8-UV5 lac)</font></li> |
<li><font color="#FF3333">Restriction digestion with [E/S]</font></li> | <li><font color="#FF3333">Restriction digestion with [E/S]</font></li> | ||
<li><font color="#FF3333">Gel purification</font></li> | <li><font color="#FF3333">Gel purification</font></li> | ||
− | <li><font color="#FF3333">Ligation of the [E/S] digested lacIq into [E/S] digested pSB1C3</font></li> | + | <li><font color="#FF3333">Ligation of the [E/S] digested <i>lacIq</i> into [E/S] digested pSB1C3</font></li> |
− | <br><li><font color="#0099FF">Ligation of the [E/S] digested lacIq and [X/P] digested λLysis_Sm(o_c_c) into [E/P] digested pSB1C3</font></li></td> | + | <br><li><font color="#0099FF">Ligation of the [E/S] digested <i>lacIq</i> and [X/P] digested λLysis_Sm(o_c_c) into [E/P] digested pSB1C3</font></li></td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
Line 833: | Line 833: | ||
<td></td> | <td></td> | ||
<td></td> | <td></td> | ||
− | <td><li><font color="#FF3333"> The Aug 4 transformed pSB1C3-lacIq showed no growth</font></li> | + | <td><li><font color="#FF3333"> The Aug 4 transformed pSB1C3-<i>lacIq</i> showed no growth</font></li> |
<li><font color="#FF9900">Colony PCR on Aug 4 transformed λLysis_Sm(c_c_c) clones. The clones contained inserts of the correct size</font></li> | <li><font color="#FF9900">Colony PCR on Aug 4 transformed λLysis_Sm(c_c_c) clones. The clones contained inserts of the correct size</font></li> | ||
− | <li><font color="#0099FF">Colony PCR on Aug 4 transformed lacIq-λLysis_Sm(o_c_c) clones. The clones contained inserts of the wrong sizes</font></li> | + | <li><font color="#0099FF">Colony PCR on Aug 4 transformed <i>lacIq</i>-λLysis_Sm(o_c_c) clones. The clones contained inserts of the wrong sizes</font></li> |
</td> | </td> | ||
</tr> | </tr> | ||
Line 846: | Line 846: | ||
<td><li><font color="#0099FF">Restriction digestion of λLysis_Sm(o_c_c) with [E/X]</li> | <td><li><font color="#0099FF">Restriction digestion of λLysis_Sm(o_c_c) with [E/X]</li> | ||
<li>Gel purification</font></li> | <li>Gel purification</font></li> | ||
− | <li><font color="#FF9900">Redo the ligation of [E/S] digested lacIq into the [E/X] digested pSB1C3-λLysis_Sm(c_c_c)</font></li> | + | <li><font color="#FF9900">Redo the ligation of [E/S] digested <i>lacIq</i> into the [E/X] digested pSB1C3-λLysis_Sm(c_c_c)</font></li> |
− | <li><font color="#FF3333"> Redo the ligation of the [E/S] digested lacIq into [E/S] digested pSB1C3</font></li></td> | + | <li><font color="#FF3333"> Redo the ligation of the [E/S] digested <i>lacIq</i> into [E/S] digested pSB1C3</font></li></td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
Line 854: | Line 854: | ||
<td></td> | <td></td> | ||
<td></td> | <td></td> | ||
− | <td><li><font color="#0099FF"> Redo the ligation of the [E/S] digested lacIq into the [E/X] digested pSB1C3-λLysis_Sm(o_c_c)</li><li>Transformation</font></li></td> | + | <td><li><font color="#0099FF"> Redo the ligation of the [E/S] digested <i>lacIq</i> into the [E/X] digested pSB1C3-λLysis_Sm(o_c_c)</li><li>Transformation</font></li></td> |
</tr> | </tr> | ||
Line 869: | Line 869: | ||
<td></td> | <td></td> | ||
<td></td> | <td></td> | ||
− | <td><li>Colony PCR on Aug 7 transformed cells. <font color="#FF3333">The lacIq clones contained inserts of the correct size.</font> <font color="#0099FF"> The lacIq-λLysis_Sm(o_c_c) clones showed no bands</font></li> | + | <td><li>Colony PCR on Aug 7 transformed cells. <font color="#FF3333">The <i>lacIq</i> clones contained inserts of the correct size.</font> <font color="#0099FF"> The <i>lacIq</i>-λLysis_Sm(o_c_c) clones showed no bands</font></li> |
<li><font color="#FF9900">Redo the restriction digestion of λLysis_Sm(c_c_c) with [E/X]</li> | <li><font color="#FF9900">Redo the restriction digestion of λLysis_Sm(c_c_c) with [E/X]</li> | ||
<li>Gel purification</li> | <li>Gel purification</li> | ||
− | <li>Ligation of the [E/S] digested lacIq into the [E/X] digested pSB1C3-λLysis_Sm(c_c_c)</font></li></td> | + | <li>Ligation of the [E/S] digested <i>lacIq</i> into the [E/X] digested pSB1C3-λLysis_Sm(c_c_c)</font></li></td> |
</table> | </table> | ||
Line 890: | Line 890: | ||
<td></td> | <td></td> | ||
<td></td> | <td></td> | ||
− | <td><li><font color="#0099FF">Redo the colony PCR on Aug 7 transformed lacIq-λLysis_Sm(o_c_c) cells. The clones contained inserts of the wrong sizes</font></li> <li><font color="#FF9900">Transformation of Aug 9 ligation product pSB1C3-lacIq-λLysis_Sm(c_c_c)</font></li></td> | + | <td><li><font color="#0099FF">Redo the colony PCR on Aug 7 transformed <i>lacIq</i>-λLysis_Sm(o_c_c) cells. The clones contained inserts of the wrong sizes</font></li> <li><font color="#FF9900">Transformation of Aug 9 ligation product pSB1C3-<i>lacIq</i>-λLysis_Sm(c_c_c)</font></li></td> |
</table> | </table> |
Latest revision as of 03:43, 20 August 2015
Keys to the table:
ori: original gene sequence, without codon optimization
co: codon optimized
λ Lysis(o_o_o): SλWT(ori)-Rλ(ori)-Rzλ(ori)
λLysis(c_c_c): SλWT(co)-Rλ(co)-Rzλ(co)
λLysis_Sm(o_c_c): Sλmut(ori)-Rλ(co)-Rzλ(co)
λLysis_Sm(c_c_c): Sλmut(co)-Rλ(co)-Rzλ(co)
[x/y]: restriction sites digested: E, EcoRI; X, XbaI; S, SpeI; P, PstI
Gene name or plasmid name [x/y]: the gene fragment or plasmid flanked with the restriction sites x and y at the ends
-A ribosome binding site (RBS) is linked upstream of each gene
-Coloring refers to sub-tasks in that particular group
-The cells used for transformation were competent JM109 E. coli cells
Week 4 : June 8 – 12 | ||||
---|---|---|---|---|
Date | Group 1 | Group 2 | Group 3 | Group 4 |
Monday 8 | ||||
Tuesday 9 | ||||
Wednesday 10 | ||||
Thursday 11 | ||||
Friday 12 |
Week 5 : June 15 – 19 | ||||
---|---|---|---|---|
Date | Group 1 | Group 2 | Group 3 | Group 4 |
Monday 15 | ||||
Tuesday 16 | ||||
Wednesday 17 | ||||
Thursday 18 | ||||
Friday 19 |
Week 6 : June 22 – 26 | ||||
---|---|---|---|---|
Date | Group 1 | Group 2 | Group 3 | Group 4 |
Monday 22 | ||||
Tuesday 23 | ||||
Wednesday 24 | ||||
Thursday 25 | ||||
Friday 26 |
Week 7 : June 29 – July 3 | ||||
---|---|---|---|---|
Date | Group 1 | Group 2 | Group 3 | Group 4 |
Monday 29 | ||||
Tuesday 30 | ||||
Wednesday 1 | ||||
Thursday 2 | ||||
Friday 3 | [1] |
Week 8 : July 6 – July 10 | ||||
---|---|---|---|---|
Date | Group 1 | Group 2 | Group 3 | Group 4 |
Monday 6 | ||||
Tuesday 7 | ||||
Wednesday 8 | ||||
Thursday 9 | ||||
Friday 10 |
Week 9 : July 13 – July 18 | ||||
---|---|---|---|---|
Date | Group 1 | Group 2 | Group 3 | Group 4 |
Monday 13 | ||||
Tuesday 14 | ||||
Wednesday 15 | ||||
Thursday 16 | ||||
Friday 17 | ||||
Saturday 18 |
DNA sequencing results later revealed that the genes amplified from the BioBrick, BBa_K124017, did not contain the desired sequences. Genomic DNA from E. coli BL21(DE3) will be used as template (starting from week 10) to create new λ(ori) related BioBricks. Gene sequences of the λ(co) clones were correct.
Week 10 : July 20 – July 25 | ||||
---|---|---|---|---|
Date | Group 1 | Group 2 | Group 3 | Group 4 |
Monday 20 | ||||
Tuesday 21 | ||||
Wednesday 22 | ||||
Thursday 23 | ||||
Friday 24 | ||||
Saturday 25 |
Week 11 : July 27 – August 1 | ||||
---|---|---|---|---|
Date | Group 1 | Group 2 | Group 3 | Group 4 |
Monday 27 | ||||
Tuesday 28 | ||||
Wednesday 29 | ||||
Thursday 30 | ||||
Friday 31 | Redo the restriction digestion of Ribo_Rλ(co) with [E/P] | |||
Saturday 1 |
Week 12 : August 3 – August 9 | ||||
---|---|---|---|---|
Date | Group 1 | Group 2 | Group 3 | Group 4 |
Monday 3 | ||||
Tuesday 4 | ||||
Wednesday 5 | ||||
Thursday 6 | ||||
Friday 7 | ||||
Saturday 8 | ||||
Sunday 9 |
Week 13 : August 10 – August 16 | ||||
---|---|---|---|---|
Date | Group 1 | Group 2 | Group 3 | Group 4 |
Monday 10 |