Cloning the shared components of complex DNA resources |
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| Authors: | Brookes, Anthony J. SIorach, Euan M. Morrison, Karen E. Qureshl, Safia J. Blake, Derek Davies, Kay Porteous, David J. |
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| Affiliation: | Molecular Genetics Section, MRC Human Genetics Unit, Western General Hospital, Crewe Road, Edinburgh EH4 2XU 1Neurosciences Group 2Molecular Genetics Group, Institute of Molecular Medicine, John Radctiffe Hospital, Oxford 0X3 9DU, UK |
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| Abstract: | The complex and repetitive nature of mammalian genomes limitsthe ability of conventional molecular techniques to recoversequences of interest. Here we describe a rapid and simple procedurefor the direct cloning of sequences which are coincident betweenDNA mixtures of whole genome complexity. The system, calledend ligation coincident sequence cloning (EL-CSC), can enrichcoincident DNA by greater than 106-fold and overcomes problemsassociated with repetitive elements. Applying EL-CSC to variouspaired DNA resources enables the facile cloning of both genomicmarkers and novel genes. To demonstrate the power of the methodwe have 1) selectively purified single copy sequences from acomplete genome, and II) isolated gene fragments from 260 kbof cloned genomic DNA. |
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