Excessive addition split peak formed by the non-templated nucleotide addition property of Taq DNA polymerase after PCR amplification.

ABSTRACT: Because of its non-template addition feature, Taq DNA polymerase can catalyze one or more extra nucleotides onto the 3' terminus of PCR products. An extra peak is observed at DYS391 locus after the PCR products stored for 4 days at 4°C. To explore the formation mechanism of this artifact, PCR primers and amplicon sequences of Y-STR loci are analyzed, furthermore, PCR products storage conditions and termination of PCR are discussed. The extra peak is a + 2 addition product, which we call excessive addition split peak (EASP). The most significant difference between EASP and the incomplete addition of adenine product is that the size of EASP is about one base larger than the true allele, and the EASP locates on the right side of the real allelic peak. The EASP cannot be eliminated by i

SUBMITTER: Zhou Y 

PROVIDER: S-EPMC10174434 | biostudies-literature | 2023

REPOSITORIES: biostudies-literature