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Cloning the shared components of complex DNA resources

Cloning the shared components of complex DNA resources
Cloning the shared components of complex DNA resources
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.
2011-2017
Brookes, Anthony J.
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SIorach, Euan M.
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Morrison, Karen E.
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Qureshl, Safia J.
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Blake, Derek
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Davies, Kay
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Porteous, David J.
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Brookes, Anthony J.
fdf91a93-e88b-4fb2-8e42-7fc8bb33a1e0
SIorach, Euan M.
8f764e06-e334-4c43-aaac-e1d19f61fbaf
Morrison, Karen E.
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Qureshl, Safia J.
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Blake, Derek
c887d206-13bf-44ce-9156-38a9e4e23e58
Davies, Kay
83a35a2a-5269-476a-a530-75618486c55a
Porteous, David J.
528a267b-2740-4e39-b4e2-83b34170d6c3

Brookes, Anthony J., SIorach, Euan M., Morrison, Karen E., Qureshl, Safia J., Blake, Derek, Davies, Kay and Porteous, David J. (1994) Cloning the shared components of complex DNA resources. Human Molecular Genetics, 3 (11), 2011-2017. (doi:10.1093/hmg/3.11.2011). (PMID:7874119)

Record type: Article

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.

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More information

Accepted/In Press date: 6 September 1994
Published date: 1994
Organisations: Medical Education

Identifiers

Local EPrints ID: 399552
URI: https://eprints.soton.ac.uk/id/eprint/399552
PURE UUID: 4f1d2161-4596-4dfa-9faa-a2a44a7569a8
ORCID for Karen E. Morrison: ORCID iD orcid.org/0000-0003-0216-5717

Catalogue record

Date deposited: 18 Aug 2016 13:02
Last modified: 06 Jun 2018 12:19

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Contributors

Author: Anthony J. Brookes
Author: Euan M. SIorach
Author: Safia J. Qureshl
Author: Derek Blake
Author: Kay Davies
Author: David J. Porteous

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