Cationic copolymers that enhance wild-type-specific suppression in BNA-clamp PCR and preferentially increase the T _m of fully matched complementary DNA and BNA strands.

ABSTRACT: Mutation detection is of major interest in molecular diagnostics, especially in the field of oncology. However, detection can be challenging as mutant alleles often coexist with excess copies of wild-type alleles. Bridged nucleic acid (BNA)-clamp PCR circumvents this challenge by preferentially suppressing the amplification of wild-type alleles and enriching rare mutant alleles. In this study, we screened cationic copolymers containing nonionic and anionic repeat units for their ability to (i) increase the T_m of double-stranded DNA, (ii) avoid PCR inhibition, and (iii) enhance the suppression of wild-type amplification in BNA-clamp PCR to detect the KRAS G13D mutation. The selected copolymers that met these criteria consisted of four types of amines and anionic and/or non

SUBMITTER: Tachibana A 

PROVIDER: S-EPMC9154250 | biostudies-literature | 2022

REPOSITORIES: biostudies-literature