▪ July 5
Anaerobic Promoter
☀ Yi Han & Clarice ☀
Colonies for EsaGFP grew
Colony PCR for 14 colonies
Unsuccessful-> no bands observed
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▪ 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
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▪ 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 |
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▪ 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 |
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▪ 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 |
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