NDLI: A method for difference cloning: gene amplification following subtractive hybridization.

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A method for difference cloning: gene amplification following subtractive hybridization.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Bolger, G. Wieland, I. Asouline, G. Wigler, M.
Description	Author Affiliation: Wieland I ( Cold Spring Harbor Laboratory, NY 11724.);
Abstract	We describe a procedure for genomic difference cloning, a method for isolating sequences present in one genomic DNA population ('tester') that is absent in another ('driver'). By subtractive hybridization, a large excess of driver is used to remove sequences common to a biotinylated tester, enriching the 'target' sequences that are unique to the tester. After repeated subtractive hybridization cycles, tester is separated from driver by avidin/biotin affinity chromatography, and single-stranded target is amplified by the polymerase chain reaction, rendering it double-stranded and clonable. We model two situations: the gain of sequences that result from infection with a pathogen and the loss of sequences that result from a large hemizygous deletion. We obtain 100- to 700-fold enrichment of target sequences.
ISSN	00278424
e-ISSN	10916490
Journal	Proceedings of the National Academy of Sciences of the United States of America
Issue Number	7
Volume Number	87
Language	English
Publisher	National Academy of Sciences
Publisher Date	1990-05-01
Publisher Place	United States
Access Restriction	Open
Subject Keyword	Cloning, Molecular DNA Genetics Nucleic Acid Amplification Techniques Nucleic Acid Hybridization Polymerase Chain Reaction Bacteriophage Lambda Cell Line Isolation & Purification DNA, Viral Molecular Sequence Data Oligonucleotide Probes Placenta Pregnancy Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. Multidisciplinary
Content Type	Text
Resource Type	Article
Subject	Multidisciplinary

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