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八聚体引发测序技术:脱氧核苷三磷酸补充的影响

Octamer-primed sequencing technology: effects of dNTP supplementation.

作者信息

Kraltcheva A I, Hardin S H

机构信息

Department of Biology and Biochemistry and Institute of Molecular Biology, University of Houston, Houston, TX 77204-5513, USA.

出版信息

Nucleic Acids Res. 2001 May 15;29(10):E48. doi: 10.1093/nar/29.10.e48.

Abstract

Octamer-primed sequencing is a directed DNA sequencing strategy that employs the use of a presynthesized octamer primer library. Together with electronic octamer sequencing technology (eOST) it provides a faster, less expensive way to obtain DNA sequence information and can be used as a valuable tool for gap closure in large-scale genomic sequencing. In this paper we discuss the effect of dGTP/TTP supplementation on octamer sequencing. We show that addition of 75 microM dGTP and 5 microM TTP can improve the sequencing success rate by increasing the length and accuracy of generated sequence information. We also discuss the effect of template base composition immediately downstream of the octamer primer on the outcome of octamer sequencing.

摘要

八聚体引发测序是一种定向DNA测序策略,它使用预先合成的八聚体引物文库。与电子八聚体测序技术(eOST)一起,它提供了一种更快、更便宜的获取DNA序列信息的方法,并且可以用作大规模基因组测序中填补缺口的有价值工具。在本文中,我们讨论了dGTP/TTP补充对八聚体测序的影响。我们表明,添加75微摩尔dGTP和5微摩尔TTP可以通过增加生成的序列信息的长度和准确性来提高测序成功率。我们还讨论了八聚体引物下游紧邻的模板碱基组成对八聚体测序结果的影响。

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