一种制备富含全长序列的标准化cDNA文库的方法

[A method for the preparation of normalized cDNA libraries enriched with full-length sequences].

作者信息

Zhulidov P A, Bogdanova E A, Shcheglov A S, Shagina I A, Vagner L L, Khazpekov G L, Kozhemiako V V, Luk'ianov S A, Shagin D A

出版信息

Bioorg Khim. 2005 Mar-Apr;31(2):186-94. doi: 10.1007/s11171-005-0023-7.

DOI:10.1007/s11171-005-0023-7

PMID:15889793

Abstract

We developed a new method for the preparation of normalized cDNA libraries enriched with full-length sequences. It is based on the properties of the recently characterized duplex-specific nuclease from the hepatopancreas of the Kamchatka crab. The duplex-specific nuclease is thermostable, it effectively cleaves double-stranded DNA and is inactive toward single-stranded DNA (Shagin et al., Genome Res., 2002, vol. 12, pp. 1935-1942). Our method enables the normalization of cDNA samples enriched with full-length sequences without use of laborious and ineffective stages of physical separation. The efficiency of the method was demonstrated in model experiments using cDNA samples from several human tissues.

摘要

我们开发了一种制备富含全长序列的标准化cDNA文库的新方法。该方法基于最近从堪察加蟹肝胰腺中鉴定出的双链特异性核酸酶的特性。双链特异性核酸酶具有热稳定性，能有效切割双链DNA，而对单链DNA无活性（沙金等人，《基因组研究》，2002年，第12卷，第1935 - 1942页）。我们的方法能够对富含全长序列的cDNA样本进行标准化，而无需使用费力且低效的物理分离步骤。在使用来自几种人类组织的cDNA样本进行的模型实验中证明了该方法的有效性。