840:153g:Projects/project10/2010/11/11: Difference between revisions
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(Autocreate 2010/11/11 Entry for 840:153g:Projects/project10) |
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== | ==BOOM! BAM! KAZAAM!== | ||
Tuesday, ligated PCR DNA with t-vector plasmid in combinations G+C 3.5:0.5, G+C 3.0:1.0, S+C 3.5:0.5, S+C 3.0:1.0 and H2O + 1.0. Thursday, transformed E. coli cells with our ligation combinations and plated on Xgal + Amp. This will indicate transfer of plasmid into E.coli as Amp resistance comes from the plasmid. Blue colonies on Xgal indicate failed ligation as lacZ in the lac operon is intact; white colonies indicate the plasmid transferred into E. coli and fragmented the lacZ gene. Next, we will pick colonies based on growth. | |||
Revision as of 14:31, 11 November 2010
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BOOM! BAM! KAZAAM!Tuesday, ligated PCR DNA with t-vector plasmid in combinations G+C 3.5:0.5, G+C 3.0:1.0, S+C 3.5:0.5, S+C 3.0:1.0 and H2O + 1.0. Thursday, transformed E. coli cells with our ligation combinations and plated on Xgal + Amp. This will indicate transfer of plasmid into E.coli as Amp resistance comes from the plasmid. Blue colonies on Xgal indicate failed ligation as lacZ in the lac operon is intact; white colonies indicate the plasmid transferred into E. coli and fragmented the lacZ gene. Next, we will pick colonies based on growth.
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