A Rapid, SuperSelective Method for Detection of Single Nucleotide Variants in Caenorhabditis elegans.

ABSTRACT: With the widespread use of single nucleotide variants generated through mutagenesis screens and genome editing technologies, there is pressing need for an efficient and low-cost strategy to genotype single nucleotide substitutions. We have developed a rapid and inexpensive method for detection of point mutants through optimization of SuperSelective (SS) primers for end-point PCR in Caenorhabditis elegans Each SS primer consists of a 5' "anchor" that hybridizes to the template, followed by a noncomplementary "bridge," and a "foot" corresponding to the target allele. The foot sequence is short, such that a single mismatch at the terminal 3' nucleotide destabilizes primer binding and prevents extension, enabling discrimination of different alleles. We explored how length and sequence c

SUBMITTER: Touroutine D 

PROVIDER: S-EPMC7536863 | biostudies-literature | 2020 Oct

REPOSITORIES: biostudies-literature