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 |
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▪ Aug 19
Anaerobic Promoter
☀ Chi Yan ☀
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 |
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