Difference between revisions of "Team:SPSingapore/Notebook-Week-7"

 
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<li><a href="https://2015.igem.org/Team:SPSingapore/Team">Overview</a></li>
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        <li><a href="https://2015.igem.org/Team:SPSingapore/Anaerobic Promoter">Anaerobic Promoter</a></li>
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   <li class='last'><a href = "https://2015.igem.org/Team:SPSingapore/Notebook"><span>ENTRIES</span></a>
 
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         <li><a href = "https://2015.igem.org/Team:SPSingapore/Notebook-Week-15"><span>Week 15 (30/8 - 5/9)</span></a></li>
 
         <li><a href = "https://2015.igem.org/Team:SPSingapore/Notebook-Week-15"><span>Week 15 (30/8 - 5/9)</span></a></li>
 
         <li><a href = "https://2015.igem.org/Team:SPSingapore/Notebook-Week-16"><span>Week 16 (6/9 - 12/9)</span></a></li>
 
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         <li class = 'last'><a href = "https://2015.igem.org/Team:SPSingapore/Notebook-Week-17"><span>Week 17 (13/9 - 18/9)</span></a></li>
 
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&#9642; July 6
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<span class = "black"> esa Quorum Sensing</span>
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&emsp;&emsp;
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<font style = "color:blue;font-weight:bold;font-size:11pt;">&#9728; Clarice &#9728;</font>
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Colony PCR with different colonies (15 colonies)
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<br> FP_BB_Prefix and RP_BB_Suffix
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<br> Success
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and on plate without amp
 
and on plate without amp
 
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<br><br>
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<span class = "black"> esa Quorum Sensing</span>
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&emsp;&emsp;
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<font style = "color:blue;font-weight:bold;font-size:11pt;">&#9728; Clarice &#9728;</font>
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<br><br>
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<div class = "divnbcontent">
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Colony PCR successful - inouclated colony 2,3,4,12 in 3mL LB
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PCR-FNRsynpart
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<tr><td> dH2O </td><td> 221ul </td></tr>
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<tr><td> Buffer </td><td> 80ul </td></tr>
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<tr><td> MgCl2 </td><td> 32ul </td></tr>
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<tr><td> dNTPs </td><td> 32ul </td></tr>
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<tr><td> FP_RNE_PromoterBBP </td><td> 16ul </td></tr>
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<tr><td> GFP_FNR_Prom_GFP </td><td> 16ul </td></tr>
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<tr><td> synpart FNR </td><td> 1ul </td></tr>
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<tr><td> gotaq </td><td> 2ul </td></tr>
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<span class = "black"> esa Quorum Sensing</span>
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&emsp;&emsp;
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<font style = "color:blue;font-weight:bold;font-size:11pt;">&#9728; Clarice &#9728;</font>
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<br><br>
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<div class = "divnbcontent">
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Verify minipreppped esaGFP plasmid
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<table class = "nbtable" style = "margin-left:50px;">
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<tr><td> Buffer </td><td> 2 </td></tr>
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<tr><td> EcoRI </td><td> 1 </td></tr>
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<tr><td> Pst1 </td><td> 1 </td></tr>
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<tr><td> Plasmid </td><td> 5 </td></tr>
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<tr><td> dH2O </td><td> 11 </td></tr>
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<tr><td> Total </td><td> 20 </td></tr>
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Latest revision as of 23:18, 18 September 2015


Research Notebook

Week 7 (5/7 - 11/7)

▪ July 5

Anaerobic Promoter    ☀ Yi Han & Clarice ☀

Colonies for EsaGFP grew
Colony PCR for 14 colonies
Unsuccessful-> no bands observed


▪ July 6

esa Quorum Sensing    ☀ Clarice ☀

Colony PCR with different colonies (15 colonies)
FP_BB_Prefix and RP_BB_Suffix
Success


▪ July 7

Workshop    ☀ Yi Han ☀

Trying out an idea for workshop
Trial for blue/white screen
Streak out pGFPuv and pGEMT on plate with amp added
spread 40ul of 0.1M IPTG and 30ul of 5%xgal, dried and on plate without amp


esa Quorum Sensing    ☀ Clarice ☀

Colony PCR successful - inouclated colony 2,3,4,12 in 3mL LB

PCR-FNRsynpart
dH2O 221ul
Buffer 80ul
MgCl2 32ul
dNTPs 32ul
FP_RNE_PromoterBBP 16ul
GFP_FNR_Prom_GFP 16ul
synpart FNR 1ul
gotaq 2ul


▪ July 8

Anaerobic Promoter    ☀ Yi Han ☀

Ran gel with FNR Pcr and gfp pcr
Size of pcr product was correct but gel picture was not saved
FNR Product was pcr purified
Plasmid extraction for colony 2,3,4 of esaGFP - > 2,3 were sent for sequencing and primers did not bind, chromatogram was messed up.

Fusion PCR for FNRGFP
dH2O 218ul
10X buffer 80ul
MgCl2 23ul
dNTP 32ul
Template FNR PCR (40ng) 3ul
gfp PCR (400ng) 1ul
FP_BB_Suffix 16ul
RP_BB_Suffix 16ul
gotaq 2ul


▪ July 9

Anaerobic Promoter    ☀ Chi Yan ☀

Gel was run for fusion pcr size was correct
RE digest
PstI 1.5
EcoRI 1.5
Buffer 4
Template 3
H2O 20
Total 40


▪ July 10

Anaerobic Promoter    ☀ Chi Yan ☀

Ran gel for gfp plasmid
gel purificaiton and extraction of 4kb fragment
ligation reaction (2X) 40ul
4ul Buffer
100ng of DNA vector 3.4
157.5ng insert 16ul
2ul enz
14.6ul H2O


esa Quorum Sensing    ☀ Clarice ☀

Verify minipreppped esaGFP plasmid
Buffer 2
EcoRI 1
Pst1 1
Plasmid 5
dH2O 11
Total 20