Cost-effective and robust genotyping using double-mismatch allele-specific quantitative PCR.

ABSTRACT: For a wide range of diseases, SNPs in the genome are the underlying mechanism of dysfunction. Therefore, targeted detection of these variations is of high importance for early diagnosis and (familial) screenings. While allele-specific PCR has been around for many years, its adoption for SNP genotyping or somatic mutation detection has been hampered by its low discriminating power and high costs. To tackle this, we developed a cost-effective qPCR based method, able to detect SNPs in a robust and specific manner. This study describes how to combine the basic principles of allele-specific PCR (the combination of a wild type and variant primer) with the straightforward readout of DNA-binding dye based qPCR technology. To enhance the robustness and discriminating power, an artificial mismatch i

SUBMITTER: Lefever S 

PROVIDER: S-EPMC6377641 | biostudies-literature | 2019 Feb

REPOSITORIES: biostudies-literature