Direct construction of a chromosome-specific NotI linking library from flow-sorted chromosomes

A linking library consists of genomic DNA fragments which contain a specific rare restriction enzyme site; such clones are very useful as probes in pulsed field gel electrophoresis and in mapping and cloning large regions of DNA. However, identifying those linking clones which map to a certain chrom...

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Bibliographic Details
Published in:Nucleic acids research 1989-02, Vol.17 (4), p.1665-1677
Main Authors: WALLACE, M. R, FOUNTAIN, J. W, BRERETON, A. M, COLLINS, F. S
Format: Article
Language:English
Subjects:
Biological and medical sciences
Biotechnology
Chromosome Mapping
Chromosomes, Human, Pair 17
Cloning, Molecular
Deoxyribonucleases, Type II Site-Specific
flow cytometry
Fundamental and applied biological sciences. Psychology
gene libraries
Genetic engineering
Genetic Linkage
Genetic technics
Genetic Vectors
Humans
Lymphocytes
Methods. Procedures. Technologies
Nucleic Acid Hybridization
Restriction Mapping
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Summary:A linking library consists of genomic DNA fragments which contain a specific rare restriction enzyme site; such clones are very useful as probes in pulsed field gel electrophoresis and in mapping and cloning large regions of DNA. However, identifying those linking clones which map to a certain chromosomal region can be laborious. Therefore, we have developed a straightforward procedure for constructing a linking library directly from flow-sorted chromosomes. As a test of the approach, a NotI linking library was constructed from the chromosome 17 fraction of a flow-sort of human chromosomes, using only 70 ng of DNA. Thirteen of sixteen linking clones were mapped to chromosome 17, suggesting that the library is highly enriched for this chromosome. This method should be generally applicable to other chromosomes and enzymes as well.
ISSN:0305-1048
1362-4962
DOI:10.1093/nar/17.4.1665