A smart polymer for sequence-selective binding, pulldown, and release of DNA targets.
Base Sequence
/ genetics
DNA
/ genetics
DNA, Complementary
/ genetics
DNA, Single-Stranded
/ genetics
Electrophoresis, Polyacrylamide Gel
Fractionation, Field Flow
Glucagon
/ genetics
High-Throughput Nucleotide Sequencing
/ methods
Humans
Insulin
/ genetics
Methanol
Pancreas
/ metabolism
Prealbumin
/ genetics
Stimuli Responsive Polymers
Transcriptome
Journal
Communications biology
ISSN: 2399-3642
Titre abrégé: Commun Biol
Pays: England
ID NLM: 101719179
Informations de publication
Date de publication:
10 07 2020
10 07 2020
Historique:
received:
10
04
2020
accepted:
17
06
2020
entrez:
12
7
2020
pubmed:
12
7
2020
medline:
22
6
2021
Statut:
epublish
Résumé
Selective isolation of DNA is crucial for applications in biology, bionanotechnology, clinical diagnostics and forensics. We herein report a smart methanol-responsive polymer (MeRPy) that can be programmed to bind and separate single- as well as double-stranded DNA targets. Captured targets are quickly isolated and released back into solution by denaturation (sequence-agnostic) or toehold-mediated strand displacement (sequence-selective). The latter mode allows 99.8% efficient removal of unwanted sequences and 79% recovery of highly pure target sequences. We applied MeRPy for the depletion of insulin, glucagon, and transthyretin cDNA from clinical next-generation sequencing (NGS) libraries. This step improved the data quality for low-abundance transcripts in expression profiles of pancreatic tissues. Its low cost, scalability, high stability and ease of use make MeRPy suitable for diverse applications in research and clinical laboratories, including enhancement of NGS libraries, extraction of DNA from biological samples, preparative-scale DNA isolations, and sorting of DNA-labeled non-nucleic acid targets.
Identifiants
pubmed: 32651444
doi: 10.1038/s42003-020-1082-2
pii: 10.1038/s42003-020-1082-2
pmc: PMC7351716
doi:
Substances chimiques
DNA, Complementary
0
DNA, Single-Stranded
0
Insulin
0
Prealbumin
0
Stimuli Responsive Polymers
0
DNA
9007-49-2
Glucagon
9007-92-5
Methanol
Y4S76JWI15
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
369Références
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