User:Andrew Perry/Notebook/bbGibson/2011/06/08: Difference between revisions
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[[Image:PCR_BbGibson_tests_080611.jpg|thumb|left| Agarose gel of PCR reactions amplifying standard Biobrick vectors and some parts with new bbGibson primers. Stained with EZ-vision (Amresco). Lane 1. Mass Ruler (Fermentas), 2. pSB1T3 (linear PCR template) V.f/V.r, 3. pSB1C3 (circular plasmid template) V.f/V.r, 4. I13522 (tetR-RBS-GFP-tt) A.f/ END.r, 5. I13522 (tetR-RBS-GFP-tt) A.f/ A.r, 6. I13521 (tetR-RBS-RFP) B.f/ END.r. ]] | [[Image:PCR_BbGibson_tests_080611.jpg|thumb|left| Agarose gel of PCR reactions amplifying standard Biobrick vectors and some parts with new bbGibson primers. Stained with EZ-vision (Amresco). Lane 1. Mass Ruler (Fermentas), 2. pSB1T3 (linear PCR template) V.f/V.r, 3. pSB1C3 (circular plasmid template) V.f/V.r, 4. I13522 (tetR-RBS-GFP-tt) A.f/ END.r, 5. I13522 (tetR-RBS-GFP-tt) A.f/ A.r, 6. I13521 (tetR-RBS-RFP-tt) B.f/ END.r. ]] | ||
'''Result:''' Got product of expected size for every part except I13522 with B.f/END.r primers (lane 6). Bad miniprep template DNA ? | '''Result:''' Got product of expected size for every part except I13522 with B.f/END.r primers (lane 6). Bad miniprep template DNA ? | ||
Revision as of 21:24, 18 August 2011
 bbGibson assembly
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bbGibson PCR testsThis is the first attempt at using the new bbGibson primers to amplify some standard Biobrick vectors and parts. Used my standard PCR protocol with Phusion polymerase.
 Result: Got product of expected size for every part except I13522 with B.f/END.r primers (lane 6). Bad miniprep template DNA ? Gel purified products (Sigma GenElute kit). | |
