Difference between revisions of "Team:SPSingapore/Notebook-Week-4"

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&#10097;&#10097;&#10097; Streaked out on plates with amp.
 
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&#10097;&#10097;&#10097; Wrong antibiotic for EsaR/I plasmid &rarr; streak out again
 
&#10097;&#10097;&#10097; Wrong antibiotic for EsaR/I plasmid &rarr; streak out again
 
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&#10097;&#10097;&#10097; Transformation of 13L repeated with 1ul of DNA.
 
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&#10097;&#10097;&#10097; Miniprep of inoculated bacteria for invasin plasmid, incubate sample in 37degC for 1h 20min:
 
&#10097;&#10097;&#10097; Miniprep of inoculated bacteria for invasin plasmid, incubate sample in 37degC for 1h 20min:
 
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&#10097;&#10097;&#10097; Restriction digest for EsaR with XbaI/BamHI:
 
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Revision as of 17:23, 13 September 2015


Research Notebook

Week 1 (24/5 - 30/5)

May 25

☀ 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

☀ 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/

☀ 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

☀ Yi Han ☀

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


May 29

☀ 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

☀ 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