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

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<div class="paragraph" style="text-align:left;">Type</div></div>
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<div class="paragraph" style="text-align:left;"><font size="5">lacZY plasmid</font></div></div>
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<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 E. coli cells </p>
  
 
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<br>
 
<br>
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<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 colspan="5"></td>
 +
  </tr>
 +
  <tr>
 +
    <td>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">
 
<table id="t01" border="1">
 
   <tr>
 
   <tr>
−
     <th></th>
+
     <th colspan="5">Week 2 : May 25 – 29</th>
−
    <th></th>
+
−
    <th></th>
+
 
   </tr>
 
   </tr>
 
   <tr>
 
   <tr>
−
     <td></td>
+
     <td colspan="5"></td>
−
    <td></td>
+
−
    <td></td>
+
 
   </tr>
 
   </tr>
 
   <tr>
 
   <tr>
−
     <td></td>
+
     <td>Date</td>
−
     <td></td>
+
     <td>Group 1</td>
−
     <td></td>
+
     <td>Group 2</td>
 +
    <td>Group 3</td>
 +
    <td>Group 4</td>
 
   </tr>
 
   </tr>
 
   <tr>
 
   <tr>
−
     <td></td>
+
     <td bgcolor="#FFB2D1">Monday 25</td>
−
     <td></td>
+
     <td colspan="4" bgcolor="#FFB2D1"></td>
−
     <td></td>
+
  </tr>
 +
  <tr>
 +
     <td>Tuesday 26</td>
 +
    <td><li>Transformation of May 22 ligation product into competent E. coli cells</li></td>
 +
    <td><li>Transformation of May 22 ligation product into competent E. coli cells</li></td>
 +
    <td><li>Transformation of May 22 ligation product into competent E. coli cells</li></td>
 +
    <td><li>Transformation of May 22 ligation product into competent E. coli 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>
 
   </tr>
 
</table>
 
</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 2 : June 1 – 5</th>
 +
  </tr>
 +
  <tr>
 +
    <td colspan="5"></td>
 +
  </tr>
 +
  <tr>
 +
    <td>Date</td>
 +
    <td>Group 1</td>
 +
    <td>Group 2</td>
 +
    <td>Group 3</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>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>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>
 
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Revision as of 14:31, 19 August 2015

Notebook - City University of Hong Kong 2015

Picture
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 2 : June 1 – 5
    Date Group 1 Group 2 Group 3 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
    		•