Difference between revisions of "Team:SPSingapore/Notebook-Week-2"

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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>
 
         <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 (13/8 - 17/9)</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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Revision as of 21:31, 18 September 2015


Research Notebook

Week 16 (6/9 - 12/9)

▪ May 31

Invasin + Listerolysin    esa Quorum Sensing
☀ Xin Yi ☀

Sent EsaR clone 4 and INv-4 for sequencing.


Invasin + Listerolysin    ☀ Yan Ting ☀

RE with XbaI and PstI
     RE - 2ul
     Buffer - 5ul
     DNA (4x of miniprep=74.5ng/ul) - 13.4ul
     H2O - 29.6ul
     Incubate at 37degC for 2.5h (1130-1400)

▶ Results: Gel loaded 1kb ladder, uncut, RE digested. 100V for 1h.
Plasmid doesn't have XbaI site - RE digested DNA is a linear 6kb band.


▪ June 2

Anaerobic Promoter    ☀ Xin Yi ☀

YFP and GFP transformation results - no colonies for YFP
The GFP transformation repeated with 100ng of plasmid was sucessful. 4 colonies of gfp plasmid were inoculated in 3mL LB+amp and grown overnight.


▪ June 3

Anaerobic Promoter    ☀ Yi Han ☀

Miniprep of gfp plasmids
RE digest with EcoRI and RsaI


▪ June 4

Anaerobic Promoter    ☀ Chi Yan ☀

RE digest indicated a very faint smaller band for colonies 2-4, and hence these were likely to be positive clones


▪ June 5

Invasin + Listerolysin    esa Quorum Sensing
☀ Yan Ting ☀

Inoculated 3mLS of Inv-4 and EsaR-4 plasmid carrying bacteria into 100mL LB+ appropriate Antibiotic
Cell culture-> HEK293 cells revived from freezing down appeared detached.


▪ June 6

Invasin + Listerolysin    esa Quorum Sensing    Anaerobic Promoter
☀ Yi Han ☀

Storage of bacterial glycerol stocks for Inv-4, EsaR4 in 25% glycerol
Yanting: grew HEK293T cells in T25 flask
Gel extract to clean up gfp plasmids which loading dye had accidentally been added to.