Difference between revisions of "Team:SPSingapore/Notebook-Week-6"

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▪ June 28
 
▪ June 28
 
<br><br>
 
<br><br>
−
<span class = "black" style = "background:green"> Lab Safety</span>
+
<span class = "black"> esa Quorum Sensing</span>
 
&emsp;&emsp;
 
&emsp;&emsp;
−
<font style = "color:blue;font-weight:bold;font-size:11pt;">&#9728; SPSingapore Team &#9728;</font>
+
<font style = "color:blue;font-weight:bold;font-size:11pt;">&#9728; Adrian &#9728;</font>
 
<br><br>
 
<br><br>
 
<div class = "divnbcontent">
 
<div class = "divnbcontent">
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<!----------------------new day------------------------------>
 
<!----------------------new day------------------------------>
 +
 +
<tr class = "tablenotebook"><td><div class = "paper">
 +
&#9642; July 2
 +
<br><br>
 +
<span class = "black"> esa Quorum Sensing</span>
 +
&emsp;&emsp;
 +
<font style = "color:blue;font-weight:bold;font-size:11pt;">&#9728; Clarice &#9728;</font>
 +
<br><br>
 +
<div class = "divnbcontent">
 +
 +
<span class = "nbcontent">
 +
Fusion PCR with esaRBS, GFP
 +
</span>
 +
<br>
 +
 +
<table class = "nbtable" style = "margin-left:50px;">
 +
<tr><td> H2O </td><td> 104.07ul </td></tr>
 +
<tr><td> Buffer </td><td> 40ul </td></tr>
 +
<tr><td> MgCl2 </td><td> 16ul </td></tr>
 +
<tr><td> dNTPs </td><td> 16ul </td></tr>
 +
<tr><td> FP_BB_prefix </td><td> 8ul </td></tr>
 +
<tr><td> RP_BB_prefix </td><td> 8ul </td></tr>
 +
<tr><td> 400ng esaRBS </td><td> 2.13ul </td></tr>
 +
<tr><td> 400ng esaRBSgfp </td><td> 4.8ul </td></tr>
 +
<tr><td> gotaq </td><td> 1ul </td></tr>
 +
<tr><td> total </td><td> 200ul </td></tr>
 +
</table>
 +
<br>
 +
<span>
 +
&#9654; annealing temp 50degC
 +
<br> &#9654; Gel electrophoresis: looks correct-> PCR Purification
 +
</span>
 +
<br><br>
 +
 +
<span class = "nbcontent">
 +
RE digest of esaRBS+GFP insert
 +
</span>
 +
<br>
 +
 +
<table class = "nbtable" style = "margin-left:50px;">
 +
<tr><td> Buffer </td><td> 2 </td></tr>
 +
<tr><td> EcoRI/PstI </td><td> 1/1 </td></tr>
 +
<tr><td> Template </td><td> 2ug 5ul </td></tr>
 +
<tr><td> dH2O </td><td> 11ul </td></tr>
 +
<tr><td> Total </td><td> 20ul </td></tr>
 +
</table>
 +
<br><br>
 +
 +
 +
<span class = "nbcontent">
 +
RE digest of esaRBS+GFP  vector
 +
</span>
 +
<br>
 +
 +
<table class = "nbtable" style = "margin-left:50px;">
 +
<tr><td> Buffer </td><td> 2ul </td></tr>
 +
<tr><td> EcoRI/PstI </td><td> 1/1 </td></tr>
 +
<tr><td> Template 4ul </td><td> 2ug 5ul </td></tr>
 +
<tr><td> dH2O </td><td> 12ul </td></tr>
 +
<tr><td> Total </td><td> 20ul </td></tr>
 +
 +
</table>
 +
 +
 +
 +
 +
</div>
 +
 +
<br><br>
 +
 +
</div>
 +
<div style = "border-top: 2px solid turquoise;"></div>
 +
</td>
 +
</tr>
 +
 +
 +
 +
<!----------------------------new day--------------------------------->
 
</table>
 
</table>
 
</center>
 
</center>

Revision as of 20:18, 17 September 2015


Research Notebook

Week 6 (28/6 - 4/7)

▪ June 28

esa Quorum Sensing    ☀ Adrian ☀

BBPrefix_esaRBS PCR
H2O 221ul
Buffer 80ul
MgCl1 32ul
dNTP 32ul
FP_Bioricks Prefix 16ul
ORP_esaRBS_fragsyn 16ul
synpart_BBP_esaRBS 1ul
gotaq 2ul
Total 400ul
▶ Success-> PCR purification

esaRBS_GFP_BBsuffix
H2O 237ul
Buffer 80ul
MgCl2 32ul
dNTP 32ul
RP_BB_suffix 16ul
ORP_esaRS_GFP 16ul
plac_GFPPLASMID 1ul
gotaq 2ul
▶ Failed -> troubleshooting-> new OFP_esaRBBS_GFP
    Redid 30/6 with new primers
    Success with new primers-> fusion pcr


▪ June 29

Lab Safety    ☀ SPSingapore Team ☀

Safety Inspection for our lab by OSHE. We passed with flying colours!


▪ July 2

esa Quorum Sensing    ☀ Clarice ☀

Fusion PCR with esaRBS, GFP
H2O 104.07ul
Buffer 40ul
MgCl2 16ul
dNTPs 16ul
FP_BB_prefix 8ul
RP_BB_prefix 8ul
400ng esaRBS 2.13ul
400ng esaRBSgfp 4.8ul
gotaq 1ul
total 200ul

▶ annealing temp 50degC
▶ Gel electrophoresis: looks correct-> PCR Purification

RE digest of esaRBS+GFP insert
Buffer 2
EcoRI/PstI 1/1
Template 2ug 5ul
dH2O 11ul
Total 20ul


RE digest of esaRBS+GFP vector
Buffer 2ul
EcoRI/PstI 1/1
Template 4ul 2ug 5ul
dH2O 12ul
Total 20ul