Nature Protocols 2006-01-01

Single-nucleotide polymorphisms: analysis by mass spectrometry.

Sascha Sauer, Richard Reinhardt, Hans Lehrach, Ivo G Gut

文献索引：Nat. Protoc. 1(4) , 1761-71, (2006)

全文：HTML全文

摘要

Matrix-assisted laser desorption-ionization (MALDI) mass spectrometry has evolved as a powerful method for analyzing nucleic acids. Here we provide protocols for genotyping single-nucleotide polymorphisms (SNPs) by MALDI based on PCR and primer extension to generate allele-specific products. Furthermore, we present three different approaches for sample preparation of primer-extension products before MALDI analysis and discuss their potential areas of application. The first approach, the 'GOOD' assay, is a purification-free procedure that uses DNA-modification chemistry, including alkylation of phosphorothioate linkages in the extension primers. The other two approaches use either solid-phase extraction or microarray purification for the purification of primer-extension products. Depending on the reaction steps of the various approaches, the protocols take about 6-8 hours.