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From gels to chips: “Minisequencing” primer extension for analysis of point mutations and single nucleotide polymorphisms
Author(s) -
Syvänen AnnChristine
Publication year - 1999
Publication title -
human mutation
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.981
H-Index - 162
eISSN - 1098-1004
pISSN - 1059-7794
DOI - 10.1002/(sici)1098-1004(1999)13:1<1::aid-humu1>3.0.co;2-i
Subject(s) - primer extension , biology , primer (cosmetics) , multiplex , genetics , genotyping , point mutation , snp genotyping , single nucleotide polymorphism , multiplex polymerase chain reaction , nucleotide , microbiology and biotechnology , computational biology , polymerase chain reaction , mutation , gene , genotype , chemistry , organic chemistry
In the minisequencing primer extension reaction, a DNA polymerase is used specifically to extend a primer that anneals immediately adjacent to the nucleotide position to be analyzed with a single labeled nucleoside triphospate complementary to the nucleotide at the variant site. The reaction allows highly specific detection of point mutations and single nucleotide polymorphisms (SNPs). Because all SNPs can be analyzed with high specificty at the same reaction conditions, minisequencing is a promising reaction principle for multiplex high‐throughput genotyping assays. It is also a useful tool for accurate quantitative PCR‐based analysis. This review discusses the different approaches, ranging from traditional gel‐based formats to multiplex detection on microarrays that have been developed and applied to minisequencing assays. Hum Mutat 13:1–10, 1999. © 1999 Wiley‐Liss, Inc.