Team:SPSingapore/Notebook-Week-4

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  • Week 1 (24/5 - 30/5)
  • Week 2 (31/5 - 6/6)
  • Week 3 (7/6 - 13/6)
  • Week 4 (14/6 - 20/6)
  • Week 5 (21/6 - 27/6)
  • Week 6 (28/6 - 4/7)
  • Week 7 (5/7 - 11/7)
  • Week 8 (12/7 - 18/7)
  • Week 9 (19/7 - 25/7)
  • Week 10 (26/7 - 1/8)
  • Week 11 (2/8 - 8/8)
  • Week 12 (9/8 - 15/8)
  • Week 13 (16/8 - 22/8)
  • Week 14 (23/8 - 29/8)
  • Week 15 (30/8 - 5/9)
  • Week 16 (6/9 - 12/9)
  • Week 17 (13/8 - 17/9)

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