Issue 19, 2012
From the journal:

Analyst

Endonuclease V-assisted accurate cleavage of oligonucleotide probes controlled by deoxyinosine and deoxynucleoside phosphorothioate for sequencing-by-ligation

Yanqiang Li,ab   Zhiqiang Pan,ab   Jing Tang,ac   Dan Pu,a   Pengfeng Xiao*a  and  Zuhong Lu*a  

* Corresponding authors

a State Key Laboratory of Bioelectronics, School of Biological Science and Medical Engineering, Southeast University, Nanjing, P.R. China
E-mail: xiaopf@seu.edu.cn, zhlu@seu.edu.cn
Fax: +86-25-83793779
Tel: +86-25-83793779

b Jiangsu Key Laboratory of Anesthesiology, Department of Anesthesiology, Xuzhou Medical College, Xuzhou, Jiangsu, P.R. China

c Tongling Municipal Center for Disease Control and Prevention, Tongling, Anhui, P.R. China

Abstract

Sequencing-by-ligation (SBL) is one of the next-generation sequencing methods for massive parallel sequencing. The ligated probes used in SBL should be accurately cleaved for a better ligation in the next cycle. Here, a novel kind of oligonucleotide probe that could be accurately cleaved at the given position was proposed. Deoxynucleoside phosphorothioates were introduced into the deoxyoxanosine-containing oligonucleotide probes in order to increase the cleavage accuracy of endonuclease V on double-stranded DNA templates. The results illustrated that incorporating deoxynucleoside phosphorothioates could greatly reduce the effect of the nonsynchronous sequencing primer, and the queried bases of the DNA templates were unambiguously identified with 5 cycles of sequencing ligations. Additionally, the read length can reach up to 25 bp with high accuracy. The SBL-based method is inexpensive, has high-throughput, and is easy to operate allowing massive scale-up, miniaturization and automation.

Graphical abstract: Endonuclease V-assisted accurate cleavage of oligonucleotide probes controlled by deoxyinosine and deoxynucleoside phosphorothioate for sequencing-by-ligation

About
Cited by
Related
Download options Please wait...

Article information

DOI
https://doi.org/10.1039/C2AN35595G
Article type
Communication
Submitted
07 May 2012
Accepted
23 Jul 2012
First published
24 Jul 2012

Analyst, 2012,137, 4421-4424

Permissions

Request permissions

Endonuclease V-assisted accurate cleavage of oligonucleotide probes controlled by deoxyinosine and deoxynucleoside phosphorothioate for sequencing-by-ligation

Y. Li, Z. Pan, J. Tang, D. Pu, P. Xiao and Z. Lu, Analyst, 2012, 137, 4421 DOI: 10.1039/C2AN35595G

To request permission to reproduce material from this article, please go to the Copyright Clearance Center request page.

If you are an author contributing to an RSC publication, you do not need to request permission provided correct acknowledgement is given.

If you are the author of this article, you do not need to request permission to reproduce figures and diagrams provided correct acknowledgement is given. If you want to reproduce the whole article in a third-party publication (excluding your thesis/dissertation for which permission is not required) please go to the Copyright Clearance Center request page.

Read more about how to correctly acknowledge RSC content.

Social activity

Spotlight

Advertisements