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Using indCAPS to Detect CRISPR/Cas9 Induced Mutations. | LitMetric

Using indCAPS to Detect CRISPR/Cas9 Induced Mutations.

Bio Protoc

Curriculum in Genetics and Molecular Biology, UNC Chapel Hill, Chapel Hill, USA.

Published: September 2019

AI Article Synopsis

Article Abstract

Cleaved amplified polymorphic sequences (CAPS) assays are useful tools for detecting small mutations such as single nucleotide polymorphisms (SNPs) or insertion/deletions (indels) present in an amplified DNA fragment. A mutation that disrupts or creates a restriction site will prevent cleavage by a restriction enzyme, allowing discrimination of wild-type and mutant alleles. In cases where no convenient restriction site is present, a derived Cleaved Amplified Polymorphic Sequence (dCAPS) assay can be used, where mismatches in the primer are used to create a diagnostic restriction site. No special design constraints are present for a CAPS assay, but cases where CAPS assays can be used are infrequent. A dCAPS assay can be burdensome to design by hand, but it is more broadly applicable. This protocol will describe the use of the indCAPS tool for the design of CAPS and dCAPS primers. The indCAPS tool was designed to be compatible with indel alleles, which prior tools struggled with but have increased importance since the rise of CRISPR/Cas9 mutagenesis methods.

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Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7854099PMC
http://dx.doi.org/10.21769/BioProtoc.3374DOI Listing

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