Team:SPSingapore/Notebook-Week-13

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Research Notebook

Week 13 (16/8 - 22/8) ▪ Aug 18 Anaerobic Promoter    ☀ Yi Han ☀ PCR for the clones of pNirB-gfp from B21(1-4) and dH5alpha (1-4)with pNirB-gfp-F/R primers 5ul plasmid, 5ul H2O in each tube PCR Mastermix dNTP 8ul MgCl2 20ul Taq Polymerase 2ul Buffer 40ul F/R primers 0.8ul each H2O 40ul ▪ Aug 19 Anaerobic Promoter    ☀ Yi Han ☀ Colony PCR for pEFYP plated on (17/8) PCR Master mix For pEYFP H2O 13.25ul Buffer 5ul dNTP 5ul F/R primer 5ul 2.5mM MgCl2 12.5 GoTaq 11.25ul PCR for pNirBgfp as with (18/8) with pNirB-gfp primers, Biobricks prefix and suffix primers (Melting temperature increased to 53 degrees) 3ul of plasmid in each tube PCR Master mix for pNirB-gfp primers and BB prefix/suffix primers H2O 101.25ul Buffer 45ul dNTP 9ul Forward Primer 9ul Reverse Primer 9ul 2.5mM MgCl2 22.5ul GoTaq polymerase 2.25ul