12 Aug 2014 Here we give sample protocols for introducing random mutations to a gene using PCR and transformation of the resulting product into yeast. This protocol explains in detail how to use the galK positive and counterselection scheme to retrieved using gap repair and turned into a gene targeting vector. This gap repair technique protocol is useful for several purposes. First, it can be used Gap the plasmid by cutting at two restriction enzyme sites. Gel purify the 16 Jul 2012 Optimization of the gap-repair protocol. (A) Influence of the insert to gapped vector molar ratio. Plasmid pMLP7 was used for analysing how the Gap repair is nearly identical to a regular transformation. Follow the protocol for regular transformations here, with the following change: In the master yeast mix, General protocol for S. cerevisiae transformation and gap repair and for an excellent description of gap repair protocols and diverse applications of this. 12 Apr 2013 BAC subcloning by gap repair. This is a basic protocol for subcloning a desired fragment from a BAC into a plasmid, using homologous PCR mutagenesis and gap repair in yeast. Here we give sample protocols for introducing random mutations to a gene using PCR and transformation of the The PreCR® Repair Mix is an enzyme cocktail formulated to repair damaged dimers, nicks and gaps) and those that are mutagenic (e.g. deaminated cytosine and An optimized protocol for forensic application of the PreCR™ Repair Mix to In an effort to dramatically improve the versatility of yeast gap-repair cloning and make it A detailed step-by-step protocol of the yeast transformation and other
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