Difference between revisions of "Team:SPSingapore/Notebook-Week-1"

 
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Latest revision as of 23:16, 18 September 2015


Research Notebook

Week 1 (24/5 - 30/5)

▪ May 25

Invasin + Listerolysin    esa Quorum Sensing
☀ Yi Han ☀

Received 2 bacterial stab cultures, EsaR/I plasmid from addgene (CHL) and BBa_K299812 from iGEM HQ.
Streaked out on plates with amp.


May 26

Invasin + Listerolysin    esa Quorum Sensing    Anaerobic Promoter
☀ Adrian ☀

Transformed:
      +Kit plate 1 9N Ba_K763002 chl
      +Kit plate 4 13L BBa_E0040 amp
DNa was received in powder form in plates, and resuspended in 10ul ultrapure H2O respectively. Plates were stored in -20/


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

Plates cracked in incubator as they dried from lack of humidity.
Transfer to small incubator with beaker of water for humidity no single colonies for inv plasmid -> streak again.
Wrong antibiotic for EsaR/I plasmid → streak out again


May 28

Invasin + Listerolysin    ☀ Yi Han ☀

❱❱❱ Inoculate single colony of invasin plasmid carrying bacteria in 3mL LB+amp
❱❱❱ Transformation of 13L repeated with 1ul of DNA.

May 29

Invasin + Listerolysin    ☀ Xin Yi ☀

No colonies grew for 13L on all plates → adjust incubator, make new media

Miniprep of inoculated bacteria for invasin plasmid, incubate sample in 37degC for 1h 20min:

RE of invasin plasmid RE control
Rsal 1ul Rsal 0 ul
EcoRI 1 ul EcoRI 0 ul
INv plasmid 7.5ul Inv plasmid 7.5 ul
H2O 35.5 ul H2O 37.5 ul
NEB buffer 4.5 ul Buffer 4.5 ul

❱❱❱ Repeat transformation of 13L with 1ul
May 30

esa Quorum Sensing    ☀ Xin Yi & Yi Han ☀

❱❱❱ Miniprep of EsaR/I plasmid for 4 colonies

❱❱❱ Restriction digest for EsaR with XbaI/BamHI:
RE reaction
5ul plasmid
5ul buffer
1ul XbaI
1ul BamHI
Add H2O to 50ul