Team:Arizona State/Notebook/PCRLog
From 2011.igem.org
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OverviewThis logbook is a record of the majority of our attempts to PCR amplify CRISPR-associated (Cas) ABCDE and Cas 3 of E. coli K12 MG1655. The record begins after our attempt to individually PCR amplify each of the 6 genes, which was unsuccessful. Here, three different sets of primers were used to attempt PCR amplification of the Cas genes in two sections. NotesSuggested annealing temperatures are based on NEB Tm Calculator, as called for in the NEB Phusion DNA Polymerase protocol. Desired band for CasABCDE at ~4300bp Desired band for Cas3 at ~2667bp Primer Round 1July 17, 2011CasABCDE: Touchdown PCR, Start Temp 70, -0.2 / cycle, Final Temp 64</p>
Gel Results: [[Image:]] CasABCDE: Very faint bands near target length
Primer Round 2==Primer Round 3== |