Team:SPSingapore/Notebook-Week-2

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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 16 (6/9 - 12/9) ▪ May 31 Invasin + Listerolysin    esa Quorum Sensing ☀ Xin Yi ☀ Sent EsaR clone 4 and INv-4 for sequencing. Invasin + Listerolysin    ☀ Yan Ting ☀ RE with XbaI and PstI      RE - 2ul      Buffer - 5ul      DNA (4x of miniprep=74.5ng/ul) - 13.4ul      H2O - 29.6ul      Incubate at 37degC for 2.5h (1130-1400) ▶ Results: Gel loaded 1kb ladder, uncut, RE digested. 100V for 1h. Plasmid doesn't have XbaI site - RE digested DNA is a linear 6kb band. ▪ June 2 Anaerobic Promoter    ☀ Xin Yi ☀ YFP and GFP transformation results - no colonies for YFP The GFP transformation repeated with 100ng of plasmid was sucessful. 4 colonies of gfp plasmid were inoculated in 3mL LB+amp and grown overnight. ▪ June 3 Anaerobic Promoter    ☀ Yi Han ☀ Miniprep of gfp plasmids RE digest with EcoRI and RsaI ▪ June 4 Anaerobic Promoter    ☀ Chi Yan ☀ RE digest indicated a very faint smaller band for colonies 2-4, and hence these were likely to be positive clones ▪ June 5 Invasin + Listerolysin    esa Quorum Sensing ☀ Yan Ting ☀ Inoculated 3mLS of Inv-4 and EsaR-4 plasmid carrying bacteria into 100mL LB+ appropriate Antibiotic Cell culture-> HEK293 cells revived from freezing down appeared detached. ▪ June 6 Invasin + Listerolysin    esa Quorum Sensing    Anaerobic Promoter ☀ Yi Han ☀ Storage of bacterial glycerol stocks for Inv-4, EsaR4 in 25% glycerol Yanting: grew HEK293T cells in T25 flask Gel extract to clean up gfp plasmids which loading dye had accidentally been added to.