Cloning the shared components of complex DNA resources

Anthony J. Brookes; Euan M. SIorach; Karen E. Morrison; Safia J. Qureshl; Derek Blake; Kay Davies; David J. Porteous

doi:10.1093/hmg/3.11.2011

Cloning the shared components of complex DNA resources

Anthony J. Brookes, Euan M. SIorach, Karen E. Morrison, Safia J. Qureshl, Derek Blake, Kay Davies, David J. Porteous

School of Medicine, Dentistry and Biomedical Sciences

Research output: Contribution to journal › Article › peer-review

10 Citations (Scopus)

Abstract

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 106-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 1) selectively purified single copy sequences from a complete genome, and II) isolated gene fragments from 260 kb of cloned genomic DNA.

Original language	English
Pages (from-to)	2011-2017
Number of pages	7
Journal	Human Molecular Genetics
Volume	3
Issue number	11
DOIs	https://doi.org/10.1093/hmg/3.11.2011
Publication status	Published - 1994

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10.1093/hmg/3.11.2011

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title = "Cloning the shared components of complex DNA resources",

abstract = "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 106-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 1) selectively purified single copy sequences from a complete genome, and II) isolated gene fragments from 260 kb of cloned genomic DNA.",

author = "Brookes, {Anthony J.} and SIorach, {Euan M.} and Morrison, {Karen E.} and Qureshl, {Safia J.} and Derek Blake and Kay Davies and Porteous, {David J.}",

year = "1994",

doi = "10.1093/hmg/3.11.2011",

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T1 - Cloning the shared components of complex DNA resources

AU - Brookes, Anthony J.

AU - SIorach, Euan M.

AU - Morrison, Karen E.

AU - Qureshl, Safia J.

AU - Blake, Derek

AU - Davies, Kay

AU - Porteous, David J.

PY - 1994

Y1 - 1994

N2 - 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 106-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 1) selectively purified single copy sequences from a complete genome, and II) isolated gene fragments from 260 kb of cloned genomic DNA.

AB - 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 106-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 1) selectively purified single copy sequences from a complete genome, and II) isolated gene fragments from 260 kb of cloned genomic DNA.

U2 - 10.1093/hmg/3.11.2011

DO - 10.1093/hmg/3.11.2011

M3 - Article

SN - 0964-6906

VL - 3

SP - 2011

EP - 2017

JO - Human Molecular Genetics

JF - Human Molecular Genetics

IS - 11

ER -

Cloning the shared components of complex DNA resources

Abstract

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