Team:NYC Wetware/Notebook/Week11

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8/8/2011<br/>
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Looking to clone RBS into plasmid pSB1A10 which contain RFP. Transformed pSB1A10 from initial distribution.
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Miniprep last of E. coli genes, and made glycerol stocks. Require verification.
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Successful amplification of biobrick insert using RFC primers. And first successful D. rad PCR
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Full plate of D. rad amplifications. Gel shows 6 definite successes and 6 'maybe's.
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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.
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Inoculate large cultures of pSB1C3+RFP as backbone for our screening plasmid.
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Miniprep large cultures of pSB1C3+RFP.
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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.

Latest revision as of 03:26, 28 September 2011