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./li>

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

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