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Issue 19, 2012
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Endonuclease V-assisted accurate cleavage of oligonucleotide probes controlled by deoxyinosine and deoxynucleoside phosphorothioate for sequencing-by-ligation

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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

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Publication details

The article was received on 07 May 2012, accepted on 23 Jul 2012 and first published on 24 Jul 2012


Article type: Communication
DOI: 10.1039/C2AN35595G
Analyst, 2012,137, 4421-4424

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    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

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