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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1994
Brookes, Anthony J.
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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), .
(doi:10.1093/hmg/3.11.2011).
(PMID:7874119)
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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Accepted/In Press date: 6 September 1994
Published date: 1994
Organisations:
Medical Education
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Local EPrints ID: 399552
URI: http://eprints.soton.ac.uk/id/eprint/399552
PURE UUID: 4f1d2161-4596-4dfa-9faa-a2a44a7569a8
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Date deposited: 18 Aug 2016 13:02
Last modified: 15 Mar 2024 01:55
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Contributors
Author:
Anthony J. Brookes
Author:
Euan M. SIorach
Author:
Karen E. Morrison
Author:
Safia J. Qureshl
Author:
Derek Blake
Author:
Kay Davies
Author:
David J. Porteous
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