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Nanopore-based identification of individual nucleotides for direct RNA sequencing.


ABSTRACT: We describe a label-free ribobase identification method, which uses ionic current measurement to resolve ribonucleoside monophosphates or diphosphates in ?-hemolysin protein nanopores containing amino-cyclodextrin adapters. The accuracy of base identification is further investigated through the use of a guanidino-modified adapter. On the basis of these findings, an exosequencing approach is envisioned in which a processive exoribonuclease (polynucleotide phosphorylase) presents sequentially cleaved ribonucleoside diphosphates to a nanopore.

SUBMITTER: Ayub M 

PROVIDER: S-EPMC3899427 | biostudies-literature | 2013

REPOSITORIES: biostudies-literature

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Nanopore-based identification of individual nucleotides for direct RNA sequencing.

Ayub Mariam M   Hardwick Steven W SW   Luisi Ben F BF   Bayley Hagan H  

Nano letters 20131113 12


We describe a label-free ribobase identification method, which uses ionic current measurement to resolve ribonucleoside monophosphates or diphosphates in α-hemolysin protein nanopores containing amino-cyclodextrin adapters. The accuracy of base identification is further investigated through the use of a guanidino-modified adapter. On the basis of these findings, an exosequencing approach is envisioned in which a processive exoribonuclease (polynucleotide phosphorylase) presents sequentially clea  ...[more]

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