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Team:NYC Wetware/Notebook/Week11
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Miniprep last of E. coli genes, and made glycerol stocks. AWAITING VERIFICATION.
Successful amplification of biobrick insert using RFC primers. And first successful D. rad PCR
Full plate of D. rad amplifications. Gel shows 6 definite successes and 6 'maybe's.
PCR pBAD promoter(K206000) for insertion in front of RBS and for assembly in composite promoter-RBS biobrick. Also PCR'ed preexisting composite promoter-RBS biobricks with some succes (K081005 fail, J04500 success). PCR RBS with RFC primers and with Verification primers for composite part.
Inoculate large cultures of pSB1C3+RFP as backbone for our screening plasmid.
Miniprep large cultures of pSB1C3+RFP.
Digest pSB1C3 backbone with E+P. Digest RBS plasmid with E+X to insert pBAD in front of RBS. Digest RBS from RFC and Ver PCRs with X+P as downstream part for assmbly into pSB1C3 with pBAD. Digest pBAD (K20600) with E+S for use as upstream part in RBS plasmid pSB1A2 and in composite part in pSB1C3 backbone.
