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

被引：26

作者：

Zhulidov, PA

Bogdanova, EA

Shcheglov, AS

Shagina, IA

Wagner, LL

Khazpekov, GL

Kozhemyako, VV

Lukyanov, SA

Shagin, DA

机构：

[1] Russian Acad Sci, Shemyakin Ovchinnikov Inst Bioorgan Chem, Moscow 117997, Russia

[2] ZAO Evrogen, Moscow, Russia

[3] Russian Acad Med Sci, Cardiol Res Ctr, Moscow, Russia

[4] Russian Acad Sci, Pacific Inst Bioorgan Chem, Far E Div, Vladivostok 690022, Russia

来源：

RUSSIAN JOURNAL OF BIOORGANIC CHEMISTRY | 2005年 / 31卷 / 02期

关键词：

cDNA normalization; duplex-specific nuclease; normalized cDNA libraries;

D O I：

10.1007/s11171-005-0023-7

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

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, effectively cleaves double-stranded DNA, and is inactive toward single-stranded DNA (Shagin et at., 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.

引用

页码：170 / 177

页数：8

共 23 条

[1] Normalization and subtraction: Two approaches to facilitate gene discovery [J].