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首页> 外文期刊>The Analyst: The Analytical Journal of the Royal Society of Chemistry: A Monthly International Publication Dealing with All Branches of Analytical Chemistry >Endonuclease V-assisted accurate cleavage of oligonucleotide probes controlled by deoxyinosine and deoxynucleoside phosphorothioate for sequencing-by-ligation
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Endonuclease V-assisted accurate cleavage of oligonucleotide probes controlled by deoxyinosine and deoxynucleoside phosphorothioate for sequencing-by-ligation

机译：核酸内切酶V辅助精确切割由脱氧肌苷和脱氧核苷硫代磷酸酯控制的寡核苷酸探针，以进行连接测序

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摘要

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.
机译：连接测序（SBL）是用于大规模并行测序的下一代测序方法之一。 SBL中使用的连接探针应准确切割，以在下一个循环中更好地连接。在这里，提出了一种新型的寡核苷酸探针，可以在给定的位置准确地切割。将脱氧核苷硫代磷酸酯引入含脱氧氧嘧啶的寡核苷酸探针中，以提高核酸内切酶V在双链DNA模板上的切割准确性。结果表明，掺入脱氧核苷硫代磷酸酯可大大降低非同步测序引物的作用，并通过5次测序连接明确鉴定了DNA模板的查询碱基。此外，读取长度可以达到25 bp的高精度。基于SBL的方法价格便宜，吞吐量高并且易于操作，可实现大规模放大，小型化和自动化。

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