Difference between revisions of "Team:CityU HK/Notebook"
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<div style="height: 20px; overflow: hidden; width: 100%;"></div></div> <!-- end of lacZY --> | <div style="height: 20px; overflow: hidden; width: 100%;"></div></div> <!-- end of lacZY --> | ||
+ | |||
+ | <!---------lysis 21 ---------> | ||
+ | <div id="lysis-21" class="paragraph" style="text-align:left;"><font size="5">Lysis plasmid (phage 21)</font></div></div> | ||
+ | <hr class="styled-hr" style="width:100%;"></hr> | ||
+ | |||
+ | <div><div style="height: 20px; overflow: hidden; width: 100%;"></div> | ||
+ | |||
+ | <p>Keys to the table: <br> | ||
+ | |||
+ | <b>ori</b>: original gene sequence, without codon optimization <br> | ||
+ | |||
+ | <b>co</b>: codon optimized <br> | ||
+ | |||
+ | <b>21Lysis(o_o_o):</b> <i>S21WT(ori)-Rλ(ori)-Rzλ(ori)</i><br> | ||
+ | |||
+ | <b>21Lysis(c_c_c):</b> <i>S21WT(co)-Rλ(co)-Rzλ(co)</i><br> | ||
+ | |||
+ | <b>21Lysis_Sm(o_c_c):</b> <i>S21mut(ori)-Rλ(co)-Rzλ(co)</i><br> | ||
+ | |||
+ | <b>21Lysis_Sm(c_c_c):</b> <i>S21mut(co)-Rλ(co)-Rzλ(co)</i><br> | ||
+ | |||
+ | [x/y]: restriction sites digested: E, EcoRI; X, XbaI; S, SpeI; P, PstI <br> | ||
+ | |||
+ | Gene name or plasmid name [x/y]: the gene fragment or plasmid flanked with the restriction sites x and y at the ends <br> | ||
+ | |||
+ | -A ribosome binding site (RBS) is linked upstream of each gene <br> | ||
+ | |||
+ | -Coloring refers to sub-tasks in that particular group <br> | ||
+ | |||
+ | -The cells used for transformation were competent JM109 <i>E. coli</i> cells</p> | ||
+ | |||
+ | <style> | ||
+ | table { | ||
+ | width:100%; | ||
+ | } | ||
+ | |||
+ | th, td { | ||
+ | padding: 5px; | ||
+ | text-align: left; | ||
+ | } | ||
+ | table#t01 tr:nth-child(even) { | ||
+ | background-color: #eee; | ||
+ | } | ||
+ | table#t01 tr:nth-child(odd) { | ||
+ | background-color:#fff; | ||
+ | } | ||
+ | table#t01 th { | ||
+ | background-color: black; | ||
+ | color: white; | ||
+ | } | ||
+ | </style> | ||
+ | |||
+ | <table id="t01" border="1"> | ||
+ | <tr> | ||
+ | <th colspan="5">Week 4 : June 8 – 12</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="20%">Date</td> | ||
+ | <td width="20%">Group 1</td> | ||
+ | <td width="20%">Group 2</td> | ||
+ | <td width="20%">Group 3</td> | ||
+ | <td width="20%">Group 4</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Monday 8</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Tuesday 9</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | |||
+ | <tr> | ||
+ | <td>Wednesday 10</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td><li>Extraction of the plasmid pGOv4-<i>Rz21(co)</i> by mini-prep</li><li>Restriction digestion with [E/P]</li></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Thursday 11</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td><li>Gel purification of the June 10 [E/P] digested <i>Rz21(co)</i></li><li>Ligation of the [E/P] digested <i>Rz21(co)</i> insert into [E/P] digested pSB1C3</li><li>Transformation</li></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Friday 12</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td><li>Colony PCR on <i>Rz21(co)</i> clones showed bands of the expected size</li></td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | |||
+ | <table id="t01" border="1"> | ||
+ | <tr> | ||
+ | <th colspan="5">Week 5 : June 15 – 19</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="20%">Date</td> | ||
+ | <td width="20%">Group 1</td> | ||
+ | <td width="20%">Group 2</td> | ||
+ | <td width="20%">Group 3</td> | ||
+ | <td width="20%">Group 4</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Monday 15</td> | ||
+ | <td colspan="4" bgcolor="#FFB2D1"></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Tuesday 16</td> | ||
+ | <td><li>Plasmid extraction of the pGOv4-<i>R21(co)</i></li><li>Restriction digestion with [E/P]</li></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | |||
+ | <tr> | ||
+ | <td>Wednesday 17</td> | ||
+ | <td><li>Gel purification of the [E/P] digested <i>R21(co)</i></li><li>Ligation of the [E/P] digested <i>R21(co)</i> insert into the [E/P] digested pSB1C3</li><li>Transformation</li></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Thursday 18</td> | ||
+ | <td><li>Colony PCR confirmed the presence of the <i>R21(co)</i> insert</li></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Friday 19</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | |||
+ | <table id="t01" border="1"> | ||
+ | <tr> | ||
+ | <th colspan="5">Week 6 : June 22 – 26</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="20%">Date</td> | ||
+ | <td width="20%">Group 1</td> | ||
+ | <td width="20%">Group 2</td> | ||
+ | <td width="20%">Group 3</td> | ||
+ | <td width="20%">Group 4</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Monday 22</td> | ||
+ | <td><li>Extraction of June 17 plasmid pSB1C3-<i>R21(co)</i></li></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Tuesday 23</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td><li>Extraction of June 11 plasmid pSB1C3-<i>Rz21(co)</i></li><li>Restriction analysis with [E/P]</li></td> | ||
+ | </tr> | ||
+ | |||
+ | <tr> | ||
+ | <td>Wednesday 24</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Thursday 25</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Friday 26</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td><li>PCR amplification on phage 21 wild type lysis cassette</li></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | |||
+ | <table id="t01" border="1"> | ||
+ | <tr> | ||
+ | <th colspan="5">Week 7 : June 29 – July 3</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="20%">Date</td> | ||
+ | <td width="20%">Group 1</td> | ||
+ | <td width="20%">Group 2</td> | ||
+ | <td width="20%">Group 3</td> | ||
+ | <td width="20%">Group 4</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Monday 29</td> | ||
+ | <td><li>Restriction digestion of June 22 <i>R21(co)</i> with [S/P] & Gel purification</li></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td><li>Extraction of June 11 plasmid pSB1C3-<i>Rz21(co)</i></li> <li>Restriction digestion with [X/P]</li></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Tuesday 30</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td><li>Redo the PCR amplification on phage 21 wild type lysis cassette</li></td> | ||
+ | <td><li>Redo June 29’s restriction digestion of <i>Rz21(co)</i> with [X/P]</li></td> | ||
+ | </tr> | ||
+ | |||
+ | <tr> | ||
+ | <td>Wednesday 1</td> | ||
+ | <td colspan="4" bgcolor="#FFB2D1"></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Thursday 2</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td><li>Gel purification of [X/P] digested <i>Rz21(co)</i></li> <li>Ligation of the [X/P] digested <i>Rz21(co)</i> insert into Gp1 June 29 [S/P] digested pSB1C3-<i>R21(co)</i> in [S/P]</li></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Friday 3</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td><li>Transformation of July 2 ligation product pSB1C3-<i>R21(co)-Rz21(co)</i></li></td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | |||
+ | <table id="t01" border="1"> | ||
+ | <tr> | ||
+ | <th colspan="5">Week 8 : July 6 – July 10</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="20%">Date</td> | ||
+ | <td width="20%">Group 1</td> | ||
+ | <td width="20%">Group 2</td> | ||
+ | <td width="20%">Group 3</td> | ||
+ | <td width="20%">Group 4</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Monday 6</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td><li>Colony PCR on July 3 <i>R21(co)-Rz21(co)</i> clones. No bands were observed</li></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Tuesday 7</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td><li>Extraction of June 30 plasmid pSB1C3-<i>S21WT(ori)-R21(ori)-Rz21(ori)</i> (21Lysis(o_o_o))</li> <li>Restriction digestion with [X/P]</li></td> | ||
+ | <td><li>Redo July 6's colony PCR with other colonies. No bands were observed</li></td> | ||
+ | </tr> | ||
+ | |||
+ | <tr> | ||
+ | <td>Wednesday 8</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td><li>Redo the restriction digestion of 21Lysis(o_o_o) with [X/P]</li></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Thursday 9</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td><li>Gel purification of July 8 [X/P] digested 21Lysis(o_o_o)</li></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Friday 10</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td><li>Ligation of the [X/P] digested 21Lysis(o_o_o) insert into [X/P] digested pSB1C3 vector</li> <li>Transformation</li></td> | ||
+ | <td><li>Restriction analysis on July 3 <i>R21(co)-Rz21(co)</i> clones. The analysis confirmed the presence of the <i>R21(co)-Rz21(co)</i> insert</li></td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | |||
+ | <table id="t01" border="1"> | ||
+ | <tr> | ||
+ | <th colspan="5">Week 9 : July 13 – July 18</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="20%">Date</td> | ||
+ | <td width="20%">Group 1</td> | ||
+ | <td width="20%">Group 2</td> | ||
+ | <td width="20%">Group 3</td> | ||
+ | <td width="20%">Group 4</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Monday 13</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td><li>Colony PCR on July 10 21Lysis(o_o_o) clones. No bands were observed</li></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Tuesday 14</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td><li>Restriction analysis on the 21Lysis(o_o_o) clones. The analysis confirmed the presence of the 21Lysis(o_o_o) insert</li></td> | ||
+ | <td><li>Extraction of the plasmid pSB1C3-<i>R21(co)-Rz21(co)</i></li> <li>Restriction digestion with [X/P]</li></td> | ||
+ | </tr> | ||
+ | |||
+ | <tr> | ||
+ | <td>Wednesday 15</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td><li>Gel purification of July 14 [X/P] digested <i>R21(co)-Rz21(co)</i></li></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Thursday 16</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Friday 17</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Saturday 18</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td><li><font color="#D633FF">Extraction of the plasmids pSB1C3-<i>S21mut(ori)</i> and pSB1C3-<i>S21mut(co)</i></li> <li>Restriction digestion wth [S/P]</font></li></td> | ||
+ | </table> | ||
+ | |||
+ | <table id="t01" border="1"> | ||
+ | <tr> | ||
+ | <th colspan="5">Week 10 : July 20 – July 25</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="20%">Date</td> | ||
+ | <td width="20%">Group 1</td> | ||
+ | <td width="20%">Group 2</td> | ||
+ | <td width="20%">Group 3</td> | ||
+ | <td width="20%">Group 4</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Monday 20</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td><li><font color="#D633FF">Gel purification of July 18 [S/P] digested pSB1C3-<i>S21mut(ori)</i> and pSB1C3-<i>S21mut(co)</i></font></li> <br> <li><font color="#0099FF">Ligation of July 15 [X/P] digested <i>R21(co)-Rz21(co)</i> insert into [S/P] digested pSB1C3-<i>S21mut(ori)</i></font></li> <br> <li><font color="#FF9900">Ligation of July 15 [X/P] digested <i>R21(co)-Rz21(co)</i> insert into [S/P] digested pSB1C3-<i>S21mut(co)</i></font></li></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Tuesday 21</td> | ||
+ | <td></td> | ||
+ | <td><li>Transformation of pGOv4-Ribo_<i>S21WT(co)</i>, pGOv4-Ribo_<i>R21(co)</i> and pGOv4-Ribo_<i>Rz21(co)</i></li></td> | ||
+ | <td></td> | ||
+ | <td><li><font color="#0099FF"> Transformation of July 20 ligation product pSB1C3-<i>S21mut(ori)-R21(co)-Rz21(co)</i> (21Lysis_Sm(o_c_c))</font></li> <li><font color="#FF9900">Transformation of July 20 ligation product pSB1C3-<i>S21mut(co)-R21(co)-Rz21(co)</i> (21Lysis_Sm(c_c_c))</font></li></td> | ||
+ | </tr> | ||
+ | |||
+ | <tr> | ||
+ | <td>Wednesday 22</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td><li>Redo the transformation of pGOv4-Ribo_<i>R21(co)</i></li></td> | ||
+ | <td><li><font color="#0099FF"> Colony PCR on the 21Lysis_Sm(o_c_c) clones revealed that the colonies contained inserts of the wrong sizes</font></li><br><li><font color="#FF9900">Colony PCR on the 21Lysis_Sm(c_c_c) clones showed bands of the expected size</font></li><br><li><font color="#0099FF"> Redo the restriction digestion of <i>R21(co)-Rz21(co)</i> with [S/P]</li><li>Gel purification</font></li></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Thursday 23</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td><li>Extraction of the plasmids pGOv4-Ribo_<i>S21WT(co)</i>, pGOv4-Ribo_<i>R21(co)</i> and pGOv4-Ribo_<i>Rz21(co)</i></li><li>Restriction digestion of Ribo_<i>S21WT(co)</i> with [E/P] & [E/S], Ribo_<i>R21(co)</i> with [E/P] & [X/P] and Ribo_<i>Rz21(co)</i> with [E/P] & [X/P]</li></td> | ||
+ | <td><li><font color="#0099FF">Redo the ligation of the [X/P] digested <i>R21(co)-Rz21(co)</i> insert into [S/P] digested pSB1C3-<i>S21mut(ori)</i></li><li>Transformation</font></li><li>Ligation of [X/P] digested BBa_I13504 (GFP) insert into [S/P] digested 21Lysis(o_o_o)</li></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Friday 24</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td><li><font color="#0099FF">Colony PCR on 21Lysis_Sm(o_c_c) clones showed bands of the expected size</font></li></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Saturday 25</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </table> | ||
+ | |||
+ | <table id="t01" border="1"> | ||
+ | <tr> | ||
+ | <th colspan="5">Week 11 : July 27 – August 1</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="20%">Date</td> | ||
+ | <td width="20%">Group 1</td> | ||
+ | <td width="20%">Group 2</td> | ||
+ | <td width="20%">Group 3</td> | ||
+ | <td width="20%">Group 4</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Monday 27</td> | ||
+ | <td colspan="4" bgcolor="#FFB2D1"></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Tuesday 28</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td><li>Extraction of the plasmid pGOv4-Ribo_<i>Rz21(co)</i></li><li>Redo the restriction digestion of Ribo_<i>S21WT(co)</i> and Ribo_<i>R21(co)</i> with [E/P]</li></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | |||
+ | <tr> | ||
+ | <td>Wednesday 29</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td><li>Ligation of the [E/P] digested Ribo_<i>S21WT(co)</i> insert into the [E/P] digested pSB1C3 vector [E/P] & transformation</li> <li>Ligation of the [E/P] digested Ribo_<i>R21(co)</i> insert into [E/P] digested pSB1C3</li><li>Extraction of the plasmids pGOv4-Ribo_<i>S21WT(co)</i>, pGOv4-Ribo_<i>R21(co)</i> and pGOv4-Ribo_<i>Rz21(co)</i></li><li>Restriction digestion of Ribo_<i>S21WT(co)</i> with [E/S], Ribo_<i>R21(co)</i> with [X/P] and Ribo_<i>Rz21(co)</i> with [E/P]</li> <li>Gel purification</li></td> | ||
+ | <td><li>Redo the ligation of the [X/P] digested GFP insert into the [S/P] digested pSB1C3-21Lysis(o_o_o)</li><li>Transformation</li></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Thursday 30</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td><li>Restriction digestion of Ribo_<i>R21(co)</i> with [X/P]</li></td> | ||
+ | <td><li>Colony PCR on 21Lysis(o_o_o)-GFP clones showed bands of the expected size</li></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Friday 31</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td><li><font color="#0099FF">Restriction digestion of 21Lysis_Sm(o_c_c) with [X/P]</li><li>Gel purification</font></li> <br> <li><font color="#FF9900">Redo the extraction of the plasmid pSB1C3-21Lysis_Sm(c_c_c)</li><li>Restriction digestion of 21Lysis_Sm(c_c_c) with [X/P]</li><li>Gel purification</font></li></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Saturday 1</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | |||
+ | <table id="t01" border="1"> | ||
+ | <tr> | ||
+ | <th colspan="5">Week 12 : August 3 – 8</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="20%">Date</td> | ||
+ | <td width="20%">Group 1</td> | ||
+ | <td width="20%">Group 2</td> | ||
+ | <td width="20%">Group 3</td> | ||
+ | <td width="20%">Group 4</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Monday 3</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td><li><font color="#127633">Extraction of plasmid pGOV4-<i>S21WT(co)</i>/li> | ||
+ | <li>Restriction digestion with [E/S]</li> | ||
+ | <li>Gel purification</li> | ||
+ | <li>Ligation of the [E/S] digested <i>S21WT(co)</i> and July 22 [X/P] digested <i>R21(co)-Rz21(co)</i> into July 22 [E/P] digested pSB1C3</font></li> <br> | ||
+ | <li><font color="#00CC00">Extraction of the plasmid pGOv4-<i>lacIq</i> (pLacIQ_LacI_L8-UV5 lac)</li> | ||
+ | <li>Restriction digestion with [E/S]</li> | ||
+ | <li>Gel purification</font></li> <br> | ||
+ | <li><font color="#0099FF">Ligation of the [E/S] digested <i>lacIq</i> and [X/P] digested 21Lysis_Sm(o_c_c) into [E/P] digested pSB1C3</font></li> | ||
+ | <li><font color="#FF9900">Ligation of the [E/S] digested <i>lacIq</i> promoter cassette and [X/P] digested 21Lysis_Sm(c_c_c) into [E/P] digested pSB1C3</font></li></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Tuesday 4</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td><li>Transformation of Aug 3 ligation product</li></td> | ||
+ | </tr> | ||
+ | |||
+ | <tr> | ||
+ | <td>Wednesday 5</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td><li><font color="#127633">Colony PCR on Aug 4 transformed pSB1C3-21Lysis(c_c_c). The clones contained inserts of the expected size</font></li><br><li><font color="#0099FF">Colony PCR on Aug 4 transformed pSB1C3-lacIq-21Lysis_Sm(o_c_c). The clones contained the wrong inserts</font></li><br><li><font color="#FF9900">Colony PCR on Aug 4 transformed pSB1C3-lacIq-21Lysis_Sm(c_c_c). The clones contained the wrong inserts</font></li></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Thursday 6</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td><li><font color="#127633">Restriction digestion of 21Lysis(c_c_c) with [E/X]</li> | ||
+ | <li>Gel purification</li> | ||
+ | <li>Ligation of the [E/S] digested lacIq into the [E/X] digested pSB1C3-21Lysis(c_c_c)</font></li> | ||
+ | <li>Restriction digestion of <font color="#0099FF">21Lysis_Sm(o_c_c)</font> and <font color="#FF9900">21Lysis_Sm(c_c_c)</font> with [E/X]</li> | ||
+ | <li>Gel purification</li> | ||
+ | <li>Redo the ligation of the [E/S] digested lacIq into the [E/X] digested pSB1C3-<font color="#0099FF">21Lysis_Sm(o_c_c)</font> and pSB1C3-<font color="#FF9900">21Lysis_Sm(c_c_c)</font> respectively</li></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Friday 7</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td><li>Transformation of Aug 6 ligation product</li> <br> <li><font color="#127633">Redo the restriction digestion with [E/S] on the plasmid pGOv4-<i>S21WT(co)</i></li><li>Gel purification</font></li></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Saturday 8</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td><li>* Colony PCR on Aug 7 transformed cells. The clones <font color="#127633">lacIq-21Lysis(c_c_c)</font>, <font color="#0099FF">lacIq-21Lysis_Sm(o_c_c)</font> and <font color="#FF9900">lacIq-21Lysis_Sm(c_c_c)</font> all contained inserts of the expected sizes</li><br><li><font color="#127633">Ligation of the [E/S] digested <i>S21WT(co)</i> into the [E/S] digested pSB1C3</li><li>Transformation</font></li></td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | |||
+ | <table id="t01" border="1"> | ||
+ | <tr> | ||
+ | <th colspan="5">Week 13 : August 10 – 16</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="20%">Date</td> | ||
+ | <td width="20%">Group 1</td> | ||
+ | <td width="20%">Group 2</td> | ||
+ | <td width="20%">Group 3</td> | ||
+ | <td width="20%">Group 4</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Monday 10</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td><li><font color="#127633">Colony PCR on Aug 8 transformed cells. The <i>S21WT(co)</i> clones contained inset of the expected size</li></td> | ||
+ | </tr> | ||
+ | </table> <!-- end of lysis 21 --> | ||
+ | |||
Revision as of 14:59, 15 September 2015