Team:SPSingapore/Notebook-Week-1

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• NOTEBOOK
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• PROTOCOL
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• ENTRIES
  • 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 (13/9 - 18/9)

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