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Genotyping short tandem repeats using flow injection and electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry
Author(s) -
Hannis James C.,
Muddiman David C.
Publication year - 2001
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.234
Subject(s) - amplicon , chemistry , genotyping , electrospray ionization , polymerase chain reaction , tandem mass spectrometry , microbiology and biotechnology , mass spectrometry , microsatellite , chromatography , genotype , biology , gene , allele , biochemistry
Characterizing polymerase chain reaction (PCR) amplicons has been accomplished for the first time using flow injection analysis coupled to electrospray ionization mass spectrometry (ESI‐MS). The PCR amplicons were amplified at the human tyrosine hydroxylase short tandem repeat locus from an individual homozygotic for the 9.3 allele. One product was amplified using Pfu polymerase and yielded a blunt‐ended amplicon of 82 base‐pairs (bp) in length. The second PCR product was amplified using Taq polymerase that resulted in an amplicon with cohesive termini of 82 bp plus either mono‐ or diadenylation. The two PCR amplicons were alternatively injected using a 0.5‐µL loop at 2 µM for the Pfu amplicon and 1 µM for the Taq amplicon with a flow rate of 200 nL/min during data acquisition. Both PCR amplicons were accurately identified using mass measurements illustrating the compatibility of ESI‐MS for genotyping short tandem repeat sequences and the potential for high‐throughput genotyping of large PCR amplicons. Copyright © 2001 John Wiley & Sons, Ltd.