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Automated discrimination of polymerase chain reaction products with closely related sequences by software-based detection of characteristic peaks in product ion spectra.

作者信息

Muhammad Warees T, Tabb David L, Fox Karen F, Fox Alvin

机构信息

Department of Pathology and Microbiology, University of South Carolina School of Medicine, Columbia, SC 29209, USA.

出版信息

Rapid Commun Mass Spectrom. 2003;17(24):2755-62. doi: 10.1002/rcm.1262.

Abstract

A computer-based method is described for automated detection of peaks in product ion spectra that allows discrimination of structurally related polymerase chain reaction (PCR) products. PCR products of K-ras mutants having single nucleotide substitutions and isomeric sequence changes in positions 1 and 2 of codon 12 (e.g. TGT and GTT) were used as a model system. SpecDiff, a tool for differentiating pairs of mass spectra by identifying peaks that either differ in relative intensity between spectra or only appear in one of a pair of spectra, was created to help automate detection. This program was demonstrated to have great utility in detection of mutations and could also be useful as a general tool for differentiating other molecules of closely related structure.

摘要

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