Robust Sequence Design Space for the Isothermal Exponential Amplification of Short Oligonucleotides.
enzymes
isothermal amplification
molecular evolution
nucleic acids
oligonucleotides
Journal
Small (Weinheim an der Bergstrasse, Germany)
ISSN: 1613-6829
Titre abrégé: Small
Pays: Germany
ID NLM: 101235338
Informations de publication
Date de publication:
24 Aug 2024
24 Aug 2024
Historique:
revised:
04
08
2024
received:
27
06
2024
medline:
24
8
2024
pubmed:
24
8
2024
entrez:
24
8
2024
Statut:
aheadofprint
Résumé
Advances in isothermal amplification techniques have accelerated development in biosensing applications and the design of complex molecular devices. The exponential amplification reaction technique, or EXPAR, is uniquely positioned to process molecular information from short oligonucleotide strands (≈10 nucleotides length) typically encountered in molecular computing or microRNA detection. Despite its conceptual simplicity (requiring only a template strand and two enzymes), the issue of nonspecific background amplification has hindered broader adoption. In this work, a new system configuration is established at 37 °C to achieve significantly improved performance. Critical sequence motifs responsible for the excellent signal-to-background profile are identified and generalized as a universal adapter design framework. Orthogonal template sequences generated from the framework are implemented for a triplex reaction and successfully evaluated mixtures of multiple-target inputs in a single-step, one-pot format without the need for exogenous agents.
Identifiants
pubmed: 39180448
doi: 10.1002/smll.202405250
doi:
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
e2405250Subventions
Organisme : Singapore Ministry of Education Academic Research Fund Tier 1
ID : A-0009534-01-00
Organisme : Singapore Ministry of Education Academic Research Fund Tier 2
ID : MOE-T2EP50120-0018
Informations de copyright
© 2024 The Author(s). Small published by Wiley‐VCH GmbH.
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