Monday, September 29, 2014

Methods for genotyping #CRISPR -generated mutations/edits: interesting alternatives.

Here are two papers with nice methods that are alternatives to the commonly used heteroduplex nuclease assays, e.g. Surveyor/Cel/T7E1.    First, a very new paper showing that simple PAGE gels can be pretty good for this.   Second, a paper from January that describes an in vitro method to use Cas9 itself + guide RNA to test PCR products for changes in "cuttability" following mutagenesis/editing.    

The second paper gets around the problem of what if there are homozygous novel variants in some samples, as these are insensitive to simple heteroduplex assays unless you mix them with known wild type products first - which can be tedious and seems sort of wasteful to me personally.

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