Team:SPSingapore/Notebook-Week-7

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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 7 (5/7 - 11/7) ▪ July 5 Anaerobic Promoter    ☀ Yi Han & Clarice ☀ Colonies for EsaGFP grew Colony PCR for 14 colonies Unsuccessful-> no bands observed ▪ July 6 esa Quorum Sensing    ☀ Clarice ☀ Colony PCR with different colonies (15 colonies) FP_BB_Prefix and RP_BB_Suffix Success ▪ July 7 Workshop    ☀ Yi Han ☀ Trying out an idea for workshop Trial for blue/white screen Streak out pGFPuv and pGEMT on plate with amp added spread 40ul of 0.1M IPTG and 30ul of 5%xgal, dried and on plate without amp esa Quorum Sensing    ☀ Clarice ☀ Colony PCR successful - inouclated colony 2,3,4,12 in 3mL LB PCR-FNRsynpart dH2O 221ul Buffer 80ul MgCl2 32ul dNTPs 32ul FP_RNE_PromoterBBP 16ul GFP_FNR_Prom_GFP 16ul synpart FNR 1ul gotaq 2ul ▪ July 8 Anaerobic Promoter    ☀ Yi Han ☀ Ran gel with FNR Pcr and gfp pcr Size of pcr product was correct but gel picture was not saved FNR Product was pcr purified Plasmid extraction for colony 2,3,4 of esaGFP - > 2,3 were sent for sequencing and primers did not bind, chromatogram was messed up. Fusion PCR for FNRGFP dH2O 218ul 10X buffer 80ul MgCl2 23ul dNTP 32ul Template FNR PCR (40ng) 3ul gfp PCR (400ng) 1ul FP_BB_Suffix 16ul RP_BB_Suffix 16ul gotaq 2ul ▪ July 9 Anaerobic Promoter    ☀ Chi Yan ☀ Gel was run for fusion pcr size was correct RE digest PstI 1.5 EcoRI 1.5 Buffer 4 Template 3 H2O 20 Total 40 ▪ July 10 Anaerobic Promoter    ☀ Chi Yan ☀ Ran gel for gfp plasmid gel purificaiton and extraction of 4kb fragment ligation reaction (2X) 40ul 4ul Buffer 100ng of DNA vector 3.4 157.5ng insert 16ul 2ul enz 14.6ul H2O esa Quorum Sensing    ☀ Clarice ☀ Verify minipreppped esaGFP plasmid Buffer 2 EcoRI 1 Pst1 1 Plasmid 5 dH2O 11 Total 20