Development of an optimization pipeline of asymmetric PCR towards the generation of DNA aptamers: a guide for beginners.
Asymmetric PCR
Biotin-streptavidin-based separation
Lambda exonuclease digestion
ssDNA
Journal
World journal of microbiology & biotechnology
ISSN: 1573-0972
Titre abrégé: World J Microbiol Biotechnol
Pays: Germany
ID NLM: 9012472
Informations de publication
Date de publication:
06 Jan 2022
06 Jan 2022
Historique:
received:
05
10
2021
accepted:
11
12
2021
entrez:
6
1
2022
pubmed:
7
1
2022
medline:
27
1
2022
Statut:
epublish
Résumé
Asymmetric PCR is one of the most utilized strategies in ssDNA generation towards DNA aptamer generation due to its low cost, robustness and the low amount of starting template. Despite its advantages, careful optimization of the asymmetric PCR is still warranted to optimize the yield of ssDNA. In this present study, we have developed an extensive optimization pipeline that involves the optimization of symmetric PCR initially followed by the optimization of asymmetric PCR. In the asymmetric PCR, optimization of primer amounts/ratios, PCR cycles, annealing temperatures, template concentrations, Mg
Identifiants
pubmed: 34989899
doi: 10.1007/s11274-021-03209-w
pii: 10.1007/s11274-021-03209-w
doi:
Substances chimiques
Aptamers, Nucleotide
0
DNA, Single-Stranded
0
Biotin
6SO6U10H04
Streptavidin
9013-20-1
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
31Subventions
Organisme : Universiti Research Grant (USM)
ID : 1001.CIPPT.8011095
Informations de copyright
© 2021. The Author(s), under exclusive licence to Springer Nature B.V.
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