DNA stretching and optimization of nucleobase recognition in enzymatic nanopore sequencing.
Nanotechnology
; 26(8): 084002, 2015 Feb 27.
Article
em En
| MEDLINE
| ID: mdl-25648138
ABSTRACT
In nanopore sequencing, where single DNA strands are electrophoretically translocated through a nanopore and the resulting ionic signal is used to identify the four DNA bases, an enzyme has been used to ratchet the nucleic acid stepwise through the pore at a controlled speed. In this work, we investigated the ability of alpha-hemolysin nanopores to distinguish the four DNA bases under conditions that are compatible with the activity of DNA-handling enzymes. Our findings suggest that in immobilized strands, the applied potential exerts a force on DNA (â¼10 pN at +160 mV) that increases the distance between nucleobases by about 2.2 Å V(-1). The four nucleobases can be resolved over wide ranges of applied potentials (from +60 to +220 mV in 1 m KCl) and ionic strengths (from 200 mM KCl to 1 M KCl at +160 mV) and nucleobase recognition can be improved when the ionic strength on the side of the DNA-handling enzyme is low, while the ionic strength on the opposite side is high.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
DNA
/
Análise de Sequência de DNA
Tipo de estudo:
Prognostic_studies
Idioma:
En
Revista:
Nanotechnology
Ano de publicação:
2015
Tipo de documento:
Article
País de afiliação:
Reino Unido