Proofreading genotyping assays mediated by high fidelity exo+ DNA polymerases.
DNA polymerases with 3'-5' proofreading function mediate high fidelity DNA replication but their application for mutation detection was almost completely neglected before 1998. The obstacle facing the use of exo(+) polymerases for mutation detection could be overcome by primer-3'-termini modification, which has been tested using allele-specific primers with 3' labeling, 3' exonuclease-resistance and 3' dehydroxylation modifications. Accordingly, three new types of single nucleotide polymorphism (SNP) assays have been developed to carry out genome-wide genotyping making use of the fidelity advantage of exo(+) polymerases. Such SNP assays might also provide a novel approach for re-sequencing and de novo sequencing. These new mutation detection assays are widely adaptable to a variety of platforms, including real-time PCR, multi-well plate and microarray technologies. Application of exo(+) polymerases to genetic analysis could accelerate the pace of personalized medicine.
Zhang
Jia
J
SNP Institute, North District of the School, Nanhua University, Hengyang, Hunan, 421001, China.
Li
Kai
K
Pardinas
Jose R
JR
Sommer
Steve S
SS
Yao
Kai-Tai
KT
D017931:DNA Primers; D004259:DNA-Directed DNA Polymerase / Q000235:genetics* / Q000378:metabolism; D005838:Genotype; D006801:Humans; D009154:Mutation; D020641:Polymorphism, Single Nucleotide; D017422:Sequence Analysis, DNA / Q000379:methods
D017931:DNA Primers; D004259:DNA-Directed DNA Polymerase