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Electrospray ionization quadrupole time‐of‐flight mass spectrometry and quadrupole mass spectrometry for genotyping single nucleotide substitutions in intact polymerase chain reaction products in K‐ras and p53
Author(s) -
Muhammad Warees T.,
Fox Karen F.,
Fox Alvin,
Cotham William,
Walla Michael
Publication year - 2002
Publication title -
rapid communications in mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.528
H-Index - 136
eISSN - 1097-0231
pISSN - 0951-4198
DOI - 10.1002/rcm.859
Subject(s) - chemistry , mass spectrometry , electrospray ionization , quadrupole , quadrupole time of flight , chromatography , electrospray , nucleotide , analytical chemistry (journal) , biochemistry , physics , atomic physics , gene
Abstract Single nucleotide polymorphisms (SNPs) and mutations were genotyped for both homozygous and heterozygous PCR products of p53, a tumor suppressor gene, and K‐ras, an oncogene, using electrospray ionization (ESI) quadrupole time‐of‐flight (Q‐TOF) mass spectrometry (MS) and ESI‐quadrupole MS analysis. Mass accuracy was adequate for both instruments to detect genetic changes in homozygous PCR products, including the most difficult to distinguish (adenine [A] → thymine [T] transversion). However, for the detection of A → T shifts (9.0 Da difference) in heterozygous PCR products, the increased resolution of ESI‐Q‐TOFMS proved essential. Although, greater mass differences in heterozygotes (e.g. cytosine [C] ↔ T or guanine [G] ↔ A) can be discriminated using ESI‐quadrupole MS analysis. Copyright © 2002 John Wiley & Sons, Ltd.