CLONING THE SHARED COMPONENTS OF COMPLEX DNA RESOURCES

被引：6

作者：

BROOKES, AJ

SLORACH, EM

MORRISON, KE

QURESHI, SJ

BLAKE, D

DAVIES, K

PORTEOUS, DJ

机构：

[1] JOHN RADCLIFFE HOSP,INST MOLEC MED,NEUROSCI GRP,OXFORD OX3 9DU,ENGLAND

[2] JOHN RADCLIFFE HOSP,INST MOLEC MED,MOLEC GENET GRP,OXFORD OX3 9DU,ENGLAND

来源：

HUMAN MOLECULAR GENETICS | 1994年 / 3卷 / 11期

基金：

英国医学研究理事会; 英国惠康基金;

关键词：

D O I：

10.1093/hmg/3.11.2011

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：

The complex and repetitive nature of mammalian genomes limits the ability of conventional molecular techniques to recover sequences of interest. Here we describe a rapid and simple procedure for the direct cloning of sequences which are coincident between DNA mixtures of whole genome complexity. The system, called end ligation coincident sequence cloning (EL-CSC), can enrich coincident DNA by greater than 10(6)-fold and overcomes problems associated with repetitive elements. Applying EL-CSC to various paired DNA resources enables the facile cloning of both genomic markers and novel genes. To demonstrate the power of the method we have i) selectively purified single copy sequences from a complete genome, and ii) isolated gene fragments from 260 kb of cloned genomic DNA.

引用

页码：2011 / 2017

页数：7

共 26 条

[1] RAPID ISOLATION OF DESIRED SEQUENCES FROM LONE LINKER PCR AMPLIFIED CDNA MIXTURES - APPLICATION TO IDENTIFICATION AND RECOVERY OF EXPRESSED SEQUENCES IN CLONED GENOMIC DNA [J].