APPROACH TO GENOTYPING ERRORS CAUSED BY NONTEMPLATED NUCLEOTIDE ADDITION BY TAQ DNA-POLYMERASE

Citation
Jr. Smith et al., APPROACH TO GENOTYPING ERRORS CAUSED BY NONTEMPLATED NUCLEOTIDE ADDITION BY TAQ DNA-POLYMERASE, PCR methods and applications, 5(3), 1995, pp. 312-317
Citations number
6
Categorie Soggetti
Biothechnology & Applied Migrobiology",Biology
ISSN journal
10549803
Volume
5
Issue
3
Year of publication
1995
Pages
312 - 317
Database
ISI
SICI code
1054-9803(1995)5:3<312:ATGECB>2.0.ZU;2-2
Abstract
Thermostable DNA polymerases can catalyze nontemplated addition of a n ucleotide to the 3' end of amplification products. This presents a pot ential source of error in genotyping studies employing Taq DNA polymer ase to amplify microsatellite loci. Although the activity is marker sp ecific, experimental variation is often seen In the degree of modifica tion. Consequently, for a given microsatellite marker, an allele may b e inconsistently identified as either the unmodified or modified ampli fication product. Full automation of high-throughput genotyping has be en hampered by the need for manual editing of data because of this sou rce of allele misidentification. In this study we estimate a 1% to 3% error rate attributable to nontemplated nucleotide addition in the ABI PRISM genotyping system. We present a PCR-based strategy to minimize this source of error.