IGEM:Groningen/2009/Notebook/iGEM 2011/2011/07/05: Difference between revisions
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<br>Competent cells were prepared with the protocol of openwetware: | <br>Competent cells were prepared with the protocol of openwetware: | ||
<br>Transformation of E.coli DH5 alpha competent cells protocol: http://openwetware.org/wiki/Transforming_chemically_competent_cells | <br>Transformation of E.coli DH5 alpha competent cells protocol: http://openwetware.org/wiki/Transforming_chemically_competent_cells | ||
<br> | <br> * N.B: 10μl ligation mixture was used for each transformation | ||
Revision as of 03:30, 7 July 2011
iGEM Project name 1 | <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html> |
Entry titlePCR of HybB promotor, PcI, PlasI, LasR-LVA, cI LVA. PCR protocol: Denaturation: 94 °C 7 minutes Cycle 30× Denaturation: 94 °C 1 minute Annealing: 30 seconds (Temperature gradient: 65 °C hybB, 60 °C PcI and PlasI, 57°C LasR-LVA and cI-LVA) Extension: 2 minutes Final extension: 7 minutes Store infinite at 4 °C PCR cleanup and vector cleanup according to the High Pure PCR purification kit of Roche (LOT: 11 732 676 001 version 15.0) Check PCR cleanups at 1.5 % agarose gel: products visible, one band, right size! (During agarose gel electrophoresis nanodrop of samples: Vectors: 50 ng/μl, HybB: 70ng/μl, PcI: 65.9 ng/μl, PlasI: 42.7 ng/μl, cI-LVA: 149.3 ng/μl, LasR-LVA 127.1 ng/μl)
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