Exogenous contaminating DNA in Taq polymerases: A method to avoid false-positive results when detecting the bla TEM gene.

Autor: Masalov YK; Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, 32 Vavilova Str., Moscow 119991, Russia., Heydarov RN; Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, 32 Vavilova Str., Moscow 119991, Russia., Shashkov IA; Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, 32 Vavilova Str., Moscow 119991, Russia., Chebotar IV; National Medical Research Center for Children's Health, Ministry of Health, 2 Lomonosovsky Pros., Moscow 119991, Russia., Zasedatelev AS; Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, 32 Vavilova Str., Moscow 119991, Russia., Mayansky NA; National Medical Research Center for Children's Health, Ministry of Health, 2 Lomonosovsky Pros., Moscow 119991, Russia., Mikhailovich VM; Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, 32 Vavilova Str., Moscow 119991, Russia. Electronic address: v.mikhailovich@gmail.com.
Jazyk: angličtina
Zdroj: Journal of microbiological methods [J Microbiol Methods] 2019 May; Vol. 160, pp. 36-41. Date of Electronic Publication: 2019 Mar 20.
DOI: 10.1016/j.mimet.2019.03.018
Abstrakt: In the course of developing an assay to identify genes responsible for antibiotic resistance in gram-negative bacteria, it has been found that standard (not DNA-free) Taq DNA polymerases were contaminated with bla TEM gene fragments that varied in length and quantities. The complete bla TEM gene sequence was either absent or was detected in infinitesimal amounts. We developed an approach to avoid false-positive findings caused by contaminating bla TEM gene sequences in conventional polymerases. The method is based on selection of a target sequence to be detected within the bla TEM gene in such a way that the chosen sequence is amplified with primers incapable of amplifying contaminating DNA sequences of the polymerase.
(Copyright © 2019 Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
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