Template length, concentration and guanidine and cytosine content influence on multiple displacement amplification efficiency.
Amplification bias
Genome length
G + C content
Multiple displacement amplification
Quantitative polymerase chain reaction
Whole genome amplification
Journal
Journal of microbiological methods
ISSN: 1872-8359
Titre abrégé: J Microbiol Methods
Pays: Netherlands
ID NLM: 8306883
Informations de publication
Date de publication:
02 2021
02 2021
Historique:
received:
18
11
2020
revised:
12
01
2021
accepted:
14
01
2021
pubmed:
26
1
2021
medline:
28
9
2021
entrez:
25
1
2021
Statut:
ppublish
Résumé
Detection of low abundance human health pathogens in environmental samples is a challenge for water monitoring. This limitation can be overcome by the introduction of multiple displacement amplification (MDA) where a minute amount of genetic material can be amplified using a phi-29 DNA polymerase. However, the genetic makeup and the concentration of the polynucleotides might influence the amplification process due to inherent assay bias. Herein, a series of experiments were designed to demonstrate the effect of genome length, guanidine and cytosine content, and template concentration on the efficiency of MDA. Quantitative polymerase chain reaction (qPCR) was performed to quantify pre- and post-MDA concentrations of selected genes. Linear regression between pre- and post-MDA log gene copies L
Identifiants
pubmed: 33493489
pii: S0167-7012(21)00014-2
doi: 10.1016/j.mimet.2021.106146
pii:
doi:
Substances chimiques
DNA, Bacterial
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.
Langues
eng
Sous-ensembles de citation
IM
Pagination
106146Informations de copyright
Copyright © 2021 Elsevier B.V. All rights reserved.