Difference between revisions of "Team:CityU HK/Notebook"

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                             PCR purification
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                             LacZY plasmid
 
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                             Plasmid extraction
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                             Lysis plasmid (λ phage)
 
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                             Gel purification
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                             Lysis plasmid (Phage 21)
 
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                             Protein purification
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                             Interlab study
 
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<!-- main content starts here -->
 
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                   <center><h2>Lab Log</h2></center>
 
                   <center><h2>Lab Log</h2></center>
  
 +
<div class="wsite-spacer" style="height:50px;"></div>
 +
 +
<div id="laczy" class="paragraph" style="text-align:left;"><span id="selectionBoundary_1439072587711_8340591508895159" class="rangySelectionBoundary" style="line-height: 0; display: none;">&#65279;</span><font size="6">Notebook</font><span id="selectionBoundary_1439072587710_16871270420961082" class="rangySelectionBoundary" style="line-height: 0; display: none;">&#65279;</span></div>
 +
 +
<div><div style="height: 20px; overflow: hidden; width: 100%;"></div>
 +
<hr class="styled-hr" style="width:100%;"></hr>
 +
<div style="height: 20px; overflow: hidden; width: 100%;"></div></div>
 +
 +
<div class="paragraph" style="text-align:left;"><font size="5">lacZY plasmid</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>
 +
 +
[?/?]: Names of restriction enzyme used for digestion:; E, EcoRI; X, XbaI; S, SpeI; P, PstI <br>
 +
 +
- A ribosome binding site (RBS) is linked upstream to each gene <br>
 +
 +
- The host 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>
 +
 +
<br>
 +
<p>PCR amplification of the lacZ and lacY genes.</p>
 +
<table id="t01" border="1">
 +
  <tr>
 +
    <th colspan="5">Week 1 : May 18 – 22</th>
 +
  </tr>
 +
  <tr>
 +
    <td width="20%">Date</td>
 +
    <td>Group 1</td>
 +
    <td>Group 2</td>
 +
    <td>Group 3</td>
 +
    <td>Group 4</td>
 +
  </tr>
 +
  <tr>
 +
    <td>Monday 18</td>
 +
    <td colspan="4"><center>==introduction==</center></td>
 +
  </tr>
 +
  <tr>
 +
    <td bgcolor="#FFB2D1">Tuesday 19</td>
 +
    <td colspan="4" bgcolor="#FFB2D1"></td>
 +
  </tr>
 +
  <tr>
 +
    <td>Wednesday 20</td>
 +
    <td><li>PCR amplification of the BBa_S04055 <i>lacZ</i> gene</li></td>
 +
    <td><li>PCR amplification of the BBa_S04055 <i>lacZ</i> gene</li></td>
 +
    <td><li>PCR amplification of the BBa_S04055 <i>lacY</i> gene</li></td>
 +
    <td><li>PCR amplification of the BBa_S04055 <i>lacY</i> gene</li></td>
 +
  </tr>
 +
  <tr>
 +
    <td>Thursday 21</td>
 +
    <td><li>Gel electrophoresis of the amplicon</li> <li><i>lacZ</i> PCR product purification </li> <li>Restriction digestion with [E/P] of the vector pSB1C3 & <i>lacZ</i></li></td>
 +
    <td><li>Gel electrophoresis of the amplicon</li> <li><i>lacZ</i> PCR product purification </li> <li>Restriction digestion with [E/P] of the vector pSB1C3 & <i>lacZ</i></li></td>
 +
    <td><li>Gel electrophoresis of the amplicon</li> <li><i>lacY</i> PCR product purification </li> <li>Restriction digestion with [E/P] of the vector pSB1C3 & <i>lacY</i></li></td>
 +
    <td><li>Gel electrophoresis of the amplicon</li> <li><i>lacY</i> PCR product purification </li> <li>Restriction digestion with [E/P] of the vector pSB1C3 & <i>lacY</i></li></td>
 +
  </tr>
 +
  <tr>
 +
    <td>Friday 22</td>
 +
    <td><li>Gel purification of vector pSB1C3 [E/P] digest</li> <li>Ligation of the [E/P] digested <i>lacZ</i> insert into the [E/P] digested vector pSB1C3</li></td>
 +
    <td><li>Gel purification of vector pSB1C3 [E/P] digest</li> <li>Ligation of the [E/P] digested <i>lacZ</i> insert into the [E/P] digested vector pSB1C3</li></td>
 +
    <td><li>Gel purification of vector pSB1C3 [E/P] digest</li> <li>Ligation of the [E/P] digested <i>lacY</i> insert into the [E/P] digested vector pSB1C3</li></td>
 +
    <td><li>Gel purification of vector pSB1C3 [E/P] digest</li> <li>Ligation of the [E/P] digested <i>lacY</i> insert into the [E/P] digested vector pSB1C3</li></td>
 +
  </tr>
 +
</table>
 +
 +
<table id="t01" border="1">
 +
  <tr>
 +
    <th colspan="5">Week 2 : May 25 – 29</th>
 +
  </tr>
 +
  <tr>
 +
    <td width="20%">Date</td>
 +
    <td>Group 1</td>
 +
    <td>Group 2</td>
 +
    <td>Group 3</td>
 +
    <td>Group 4</td>
 +
  </tr>
 +
  <tr>
 +
    <td bgcolor="#FFB2D1">Monday 25</td>
 +
    <td colspan="4" bgcolor="#FFB2D1"></td>
 +
  </tr>
 +
  <tr>
 +
    <td>Tuesday 26</td>
 +
    <td><li>Transformation of May 22 ligation product into competent <i>E. coli</i> cells</li></td>
 +
    <td><li>Transformation of May 22 ligation product into competent <i>E. coli</i> cells</li></td>
 +
    <td><li>Transformation of May 22 ligation product into competent <i>E. coli</i> cells</li></td>
 +
    <td><li>Transformation of May 22 ligation product into competent <i>E. coli</i> cells</li></td>
 +
  </tr>
 +
  <tr>
 +
    <td>Wednesday 27</td>
 +
    <td><li>Colony PCR of May 26 transformed cells</li></td>
 +
    <td><li>Colony PCR of May 26 transformed cells</li></td>
 +
    <td><li>Colony PCR of May 26 transformed cells</li></td>
 +
    <td><li>Colony PCR of May 26 transformed cells</li></td>
 +
  </tr>
 +
  <tr>
 +
    <td>Thursday 28</td>
 +
    <td><li>Extraction of the plasmid pSB1C3-BBa_J23100 </li> <li>Restriction digestion with [S/P] of the vector pSB1C3-BBa_J23100</li></td>
 +
    <td><li>Extraction of the plasmid pSB1C3-BBa_J23100 </li> <li>Restriction digestion with [S/P] of the vector pSB1C3-BBa_J23100</li></td>
 +
    <td><li>Extraction of the plasmid pSB1C3-BBa_J23100 </li> <li>Restriction digestion with [S/P] of the vector pSB1C3-BBa_J23100</li></td>
 +
    <td><li>Extraction of the plasmid pSB1C3-BBa_J23100 </li> <li>Restriction digestion with [S/P] of the vector pSB1C3-BBa_J23100</li></td>
 +
  </tr>
 +
  <tr>
 +
    <td>Friday 29</td>
 +
    <td><li>Gel purification of May 28 digest</li></td>
 +
    <td><li>Gel purification of May 28 digest</li></td>
 +
    <td><li>Gel purification of May 28 digest</li></td>
 +
    <td><li>Gel purification of May 28 digest</li></td>
 +
  </tr>
 +
</table>
 +
 +
<br><br>
 +
<p>Groups 1 & 3 and Groups 2 & 4 used different cloning strategies to assemble the biobrick J23100_lacZ-lacY.</p>
 +
<br>
 +
<table id="t01" border="1">
 +
  <tr>
 +
    <th colspan="5">Week 3 : June 1 – 5</th>
 +
  </tr>
 +
  <tr>
 +
    <td width="20%">Date</td>
 +
    <td>Group 1</td>
 +
    <td>Group 3</td>
 +
    <td>Group 2</td>
 +
    <td>Group 4</td>
 +
  </tr>
 +
  <tr>
 +
    <td>Monday 1</td>
 +
    <td><li>Extraction of the plasmid pSB1C3-<i>lacZ</i> (from May 26 transformed cells)</li> <li>Restriction digestion with [X/P] of the insert <i>lacZ</i></li> <li>Gel purification</li></td>
 +
    <td><li>Extraction of the plasmid pSB1C3-<i>lacY</i> (from May 26 transformed cells)</li> <li>Restriction digestion with [X/P] of the insert <i>lacY</i></li> <li>Gel purification</li></td>
 +
    <td><li>Extraction of the plasmid pSB1C3-<i>lacZ</i> from Gp1</li> <li>Restriction digestion with [E/S] of the insert <i>lacZ</i></li> <li>Gel purification</li></td>
 +
    <td><li>Extraction of the plasmid pSB1C3-<i>lacY</i> from Gp1</li> <li>Restriction digestion with [E/S] of the insert <i>lacY</i></li> <li>Gel purification</li></td>
 +
  </tr>
 +
  <tr>
 +
    <td>Tuesday 2</td>
 +
    <td><li>Redo the work on June 1</li></td>
 +
    <td><li>Redo the work on June 1</li></td>
 +
    <td><li>Redo the work on June 1</li></td>
 +
    <td><li>Redo the work on June 1</li></td>
 +
  </tr>
 +
  <tr>
 +
    <td>Wednesday 3</td>
 +
    <td colspan="2"><li>Ligation of Gp1 [X/P] digested <i>lacZ</i> insert (from June 2) into [S/P] digested vector pSB1C3-BBa_J23100 (from May 29)</li> <li>Transformation</li></td>
 +
    <td colspan="2"><li>Ligation of Gp2 [E/S] digested <i>lacZ insert</i> (from June 2) into Gp4 [E/X] digested vector pSB1C3-<i>lacY</i>(from June 2)</li> <li>Transformation</li></td>   
 +
  </tr>
 +
  <tr>
 +
    <td>Thursday 4</td>
 +
    <td colspan="2"><li>Colony PCR on June 3 transformed cells</li></td>
 +
    <td colspan="2"><li>Colony PCR on June 3 transformed cells</li></td>
 +
  </tr>
 +
  <tr>
 +
    <td>Friday 5</td>
 +
    <td colspan="2"><li>Extraction of plasmid pSB1C3-BBa_J23100-<i>lacZ</i> (from June 3 transformed cells)</li> <li>Restriction digestion with [S/P] of the vector pSB1C3-BBa_J23100-<i>lacZ</i></li> <li>Gel purification</li></td>
 +
    <td colspan="2"><li>Extraction of plasmid pSB1C3-<i>lacZ</i>-<i>lacY</i> (from June 3 transformed cells)</li> <li>Restriction digestion with [X/P] of the insert <i>lacZ-lacY</i> <li>Gel purification</li></td>
 +
  </tr>
 +
</table>
 +
 +
<table id="t01" border="1">
 +
  <tr>
 +
    <th colspan="5">Week 4 : June 8 – 12</th>
 +
  </tr>
 +
  <tr>
 +
    <td width="20%">Date</td>
 +
    <td>Group 1</td>
 +
    <td>Group 3</td>
 +
    <td>Group 2</td>
 +
    <td>Group 4</td>
 +
  </tr>
 +
  <tr>
 +
    <td>Monday 8</td>
 +
    <td colspan="2"><li>Ligation of Gp3 [X/P] digested <i>lacY</i> insert (from June 2) into [S/P] digested vector pSB1C3-BBa_J23100-<i>lacZ</i> (from June 5)</li> <li>Transformation</li></td>
 +
    <td colspan="2"><li>Ligation of [X/P] digested <i>lacZ</i>-<i>lacY</i> insert (from June 5) into [S/P] digested vector pSB1C3-BBa_J23100 (from May 29)</li> <li>Transformation</li></td>
 +
  </tr>
 +
  <tr>
 +
    <td>Tuesday 9</td>
 +
    <td colspan="2"><li>Colony PCR of June 8 transformed cells (failed)</li></td>
 +
    <td colspan="2"><li>Colony PCR of June 8 transformed cells (failed)</li></td>
 +
  </tr>
 +
  <tr>
 +
    <td>Wednesday 10</td>
 +
    <td colspan="2"><li>Extraction of the plasmid pSB1C3-BBa_J23100-<i>lacZ</i>-<i>lacY</i> (from June 8 transformed cells)</li> <li>Restriction analysis with [E/P] for confirmation of the insert size</li></td>
 +
    <td colspan="2"><li>Extraction of the plasmid pSB1C3-BBa_J23100-<i>lacZ</i>-<i>lacY</i> (from June 8 transformed cells)</li> <li>Restriction analysis with [E/P] for confirmation of the insert size</li></td>   
 +
  </tr>
 +
</table>
 +
 +
<br>
 +
<center><u>Characterization of the lacZY plasmid</u></center>
 +
<br>
 +
 +
<table id="t01" border="1">
 +
  <tr>
 +
    <th colspan="3">Week 8 : July 6 – 10</th>
 +
  </tr>
 +
 
 +
  <tr>
 +
    <td>Date</td>
 +
    <td></td>
 +
    <td>Remarks</td>   
 +
  </tr>
 +
  <tr>
 +
    <td>Monday 6</td>
 +
    <td></td>
 +
    <td></td>
 +
  </tr>
 +
  <tr>
 +
    <td>Tuesday 7</td>
 +
    <td><li>RNA extraction from the <i>E. coli</i> cells harboring the recombinant plasmid pSB1C3-BBa_J23100- <i>lacZ</i>-<i>lacY</i></li></td>
 +
    <td>Not successful</td>
 +
  </tr>
 +
  <tr>
 +
    <td>Wednesday 8</td>
 +
    <td><li>Redo RNA extraction</li></td>
 +
    <td>Not successful</td>
 +
  </tr>
 +
  <tr>
 +
    <td>Thursday 9</td>
 +
    <td></td>
 +
    <td></td>
 +
  </tr>
 +
  <tr>
 +
    <td>Friday 10</td>
 +
    <td><li>Redo RNA extraction</li></td>
 +
    <td>Not successful</td>
 +
  </tr>
 +
 +
</table>
 +
 +
<table id="t01" border="1">
 +
  <tr>
 +
    <th colspan="3">Week 9 : July 13 – 18</th>
 +
  </tr>
 +
 
 +
  <tr>
 +
    <td>Date</td>
 +
    <td></td>
 +
    <td>Remarks</td>   
 +
  </tr>
 +
  <tr>
 +
    <td>Monday 13</td>
 +
    <td><li>Redo the RNA extraction</li></td>
 +
    <td>Successful</td>
 +
  </tr>
 +
  <tr>
 +
    <td>Tuesday 14</td>
 +
    <td><li>Extract RNA from control <i>E. coli</i> cells</li></td>
 +
    <td>Successful</td>
 +
  </tr>
 +
  <tr>
 +
    <td>Wednesday 15</td>
 +
    <td><li>Redo RNA extraction from control <i>E. coli</i> cells</li> <li>Perform RT-PCR on purified RNA from recombinant and control <i>E. coli</i></li></td>
 +
    <td>Successful</td>
 +
  </tr>
 +
  <tr>
 +
    <td>Thursday 16</td>
 +
    <td><li>Check the size of the RT-PCR product using gel electrophoresis</li></td>
 +
    <td>Successful</td>
 +
  </tr>
 +
  <tr>
 +
    <td>Friday 17</td>
 +
    <td><li>Prepare Z-buffer</li></td>
 +
    <td></td>
 +
  </tr>
 +
  <tr>
 +
    <td>Saturday 18</td>
 +
    <td><li>Perform qPCR on the control cDNA</li></td>
 +
    <td>Successful</td>
 +
 +
</table>
 +
 +
<table id="t01" border="1">
 +
  <tr>
 +
    <th colspan="3">Week 10 : July 20 – 25</th>
 +
  </tr>
 +
 
 +
  <tr>
 +
    <td>Date</td>
 +
    <td></td>
 +
    <td>Remarks</td>   
 +
  </tr>
 +
  <tr>
 +
    <td>Monday 20</td>
 +
    <td><li>Perform q-PCR on recombinant cDNA</li></td>
 +
    <td>Successful</td>
 +
  </tr>
 +
  <tr>
 +
    <td>Tuesday 21</td>
 +
    <td rowspan="4"><li>Perform ONPG assay -characterization on the expression level of LacZ protein</li></td>
 +
    <td rowspan="4"></td>
 +
  </tr>
 +
  <tr>
 +
    <td>Wednesday 22</td>
 +
   
 +
  </tr>
 +
  <tr>
 +
    <td>Thursday 23</td>
 +
   
 +
  </tr>
 +
  <tr>
 +
    <td>Friday 24</td>
 +
   
 +
  </tr>
 +
</table>
 +
 +
<div><div style="height: 20px; overflow: hidden; width: 100%;"></div>
 +
<hr class="styled-hr" style="width:100%;"></hr>
 +
<div style="height: 20px; overflow: hidden; width: 100%;"></div></div>  <!-- end of lacZY -->
  
  

Revision as of 14:55, 15 September 2015

Description - iGEM2015 wiki /* ****** Back to top button ******* */

Lab Log

Notebook
lacZY plasmid

Keys to the table:
[?/?]: Names of restriction enzyme used for digestion:; E, EcoRI; X, XbaI; S, SpeI; P, PstI
- A ribosome binding site (RBS) is linked upstream to each gene
- The host cells used for transformation were competent JM109 E. coli cells


PCR amplification of the lacZ and lacY genes.

Week 1 : May 18 – 22
Date Group 1 Group 2 Group 3 Group 4
Monday 18
==introduction==
Tuesday 19
Wednesday 20
  • PCR amplification of the BBa_S04055 lacZ gene
    		•
  • PCR amplification of the BBa_S04055 lacZ gene
    		•
  • PCR amplification of the BBa_S04055 lacY gene
    		•
  • PCR amplification of the BBa_S04055 lacY gene
    		•
    Thursday 21
  • Gel electrophoresis of the amplicon
  • lacZ PCR product purification
  • Restriction digestion with [E/P] of the vector pSB1C3 & lacZ
    		•
  • Gel electrophoresis of the amplicon
  • lacZ PCR product purification
  • Restriction digestion with [E/P] of the vector pSB1C3 & lacZ
    		•
  • Gel electrophoresis of the amplicon
  • lacY PCR product purification
  • Restriction digestion with [E/P] of the vector pSB1C3 & lacY
    		•
  • Gel electrophoresis of the amplicon
  • lacY PCR product purification
  • Restriction digestion with [E/P] of the vector pSB1C3 & lacY
    		•
    Friday 22
  • Gel purification of vector pSB1C3 [E/P] digest
  • Ligation of the [E/P] digested lacZ insert into the [E/P] digested vector pSB1C3
    		•
  • Gel purification of vector pSB1C3 [E/P] digest
  • Ligation of the [E/P] digested lacZ insert into the [E/P] digested vector pSB1C3
    		•
  • Gel purification of vector pSB1C3 [E/P] digest
  • Ligation of the [E/P] digested lacY insert into the [E/P] digested vector pSB1C3
    		•
  • Gel purification of vector pSB1C3 [E/P] digest
  • Ligation of the [E/P] digested lacY insert into the [E/P] digested vector pSB1C3
    		•
    Week 2 : May 25 – 29
    Date Group 1 Group 2 Group 3 Group 4
    Monday 25
    Tuesday 26
  • Transformation of May 22 ligation product into competent E. coli cells
    		•
  • Transformation of May 22 ligation product into competent E. coli cells
    		•
  • Transformation of May 22 ligation product into competent E. coli cells
    		•
  • Transformation of May 22 ligation product into competent E. coli cells
    		•
    Wednesday 27
  • Colony PCR of May 26 transformed cells
    		•
  • Colony PCR of May 26 transformed cells
    		•
  • Colony PCR of May 26 transformed cells
    		•
  • Colony PCR of May 26 transformed cells
    		•
    Thursday 28
  • Extraction of the plasmid pSB1C3-BBa_J23100
  • Restriction digestion with [S/P] of the vector pSB1C3-BBa_J23100
    		•
  • Extraction of the plasmid pSB1C3-BBa_J23100
  • Restriction digestion with [S/P] of the vector pSB1C3-BBa_J23100
    		•
  • Extraction of the plasmid pSB1C3-BBa_J23100
  • Restriction digestion with [S/P] of the vector pSB1C3-BBa_J23100
    		•
  • Extraction of the plasmid pSB1C3-BBa_J23100
  • Restriction digestion with [S/P] of the vector pSB1C3-BBa_J23100
    		•
    Friday 29
  • Gel purification of May 28 digest
    		•
  • Gel purification of May 28 digest
    		•
  • Gel purification of May 28 digest
    		•
  • Gel purification of May 28 digest
    		•


    Groups 1 & 3 and Groups 2 & 4 used different cloning strategies to assemble the biobrick J23100_lacZ-lacY.


    Week 3 : June 1 – 5
    Date Group 1 Group 3 Group 2 Group 4
    Monday 1
  • Extraction of the plasmid pSB1C3-lacZ (from May 26 transformed cells)
  • Restriction digestion with [X/P] of the insert lacZ
  • Gel purification
    		•
  • Extraction of the plasmid pSB1C3-lacY (from May 26 transformed cells)
  • Restriction digestion with [X/P] of the insert lacY
  • Gel purification
    		•
  • Extraction of the plasmid pSB1C3-lacZ from Gp1
  • Restriction digestion with [E/S] of the insert lacZ
  • Gel purification
    		•
  • Extraction of the plasmid pSB1C3-lacY from Gp1
  • Restriction digestion with [E/S] of the insert lacY
  • Gel purification
    		•
    Tuesday 2
  • Redo the work on June 1
    		•
  • Redo the work on June 1
    		•
  • Redo the work on June 1
    		•
  • Redo the work on June 1
    		•
    Wednesday 3
  • Ligation of Gp1 [X/P] digested lacZ insert (from June 2) into [S/P] digested vector pSB1C3-BBa_J23100 (from May 29)
  • Transformation
    		•
  • Ligation of Gp2 [E/S] digested lacZ insert (from June 2) into Gp4 [E/X] digested vector pSB1C3-lacY(from June 2)
  • Transformation
    		•
    Thursday 4
  • Colony PCR on June 3 transformed cells
    		•
  • Colony PCR on June 3 transformed cells
    		•
    Friday 5
  • Extraction of plasmid pSB1C3-BBa_J23100-lacZ (from June 3 transformed cells)
  • Restriction digestion with [S/P] of the vector pSB1C3-BBa_J23100-lacZ
  • Gel purification
    		•
  • Extraction of plasmid pSB1C3-lacZ-lacY (from June 3 transformed cells)
  • Restriction digestion with [X/P] of the insert lacZ-lacY
  • Gel purification
    		•
    Week 4 : June 8 – 12
    Date Group 1 Group 3 Group 2 Group 4
    Monday 8
  • Ligation of Gp3 [X/P] digested lacY insert (from June 2) into [S/P] digested vector pSB1C3-BBa_J23100-lacZ (from June 5)
  • Transformation
    		•
  • Ligation of [X/P] digested lacZ-lacY insert (from June 5) into [S/P] digested vector pSB1C3-BBa_J23100 (from May 29)
  • Transformation
    		•
    Tuesday 9
  • Colony PCR of June 8 transformed cells (failed)
    		•
  • Colony PCR of June 8 transformed cells (failed)
    		•
    Wednesday 10
  • Extraction of the plasmid pSB1C3-BBa_J23100-lacZ-lacY (from June 8 transformed cells)
  • Restriction analysis with [E/P] for confirmation of the insert size
    		•
  • Extraction of the plasmid pSB1C3-BBa_J23100-lacZ-lacY (from June 8 transformed cells)
  • Restriction analysis with [E/P] for confirmation of the insert size
    		•

    Characterization of the lacZY plasmid

    Week 8 : July 6 – 10
    Date Remarks
    Monday 6
    Tuesday 7
  • RNA extraction from the E. coli cells harboring the recombinant plasmid pSB1C3-BBa_J23100- lacZ-lacY
    		•  Not successful
    Wednesday 8
  • Redo RNA extraction
    		•  Not successful
    Thursday 9
    Friday 10
  • Redo RNA extraction
    		•  Not successful
    Week 9 : July 13 – 18
    Date Remarks
    Monday 13
  • Redo the RNA extraction
    		•  Successful
    Tuesday 14
  • Extract RNA from control E. coli cells
    		•  Successful
    Wednesday 15
  • Redo RNA extraction from control E. coli cells
  • Perform RT-PCR on purified RNA from recombinant and control E. coli
    		•  Successful
    Thursday 16
  • Check the size of the RT-PCR product using gel electrophoresis
    		•  Successful
    Friday 17
  • Prepare Z-buffer
    		•
    Saturday 18
  • Perform qPCR on the control cDNA
    		•  Successful
    Week 10 : July 20 – 25
    Date Remarks
    Monday 20
  • Perform q-PCR on recombinant cDNA
    		•  Successful
    Tuesday 21
  • Perform ONPG assay -characterization on the expression level of LacZ protein
    		•
    Wednesday 22
    Thursday 23
    Friday 24