Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 1990 Apr;87(7):2720-4.
doi: 10.1073/pnas.87.7.2720.

A method for difference cloning: gene amplification following subtractive hybridization

I Wieland  1 , G Bolger, G Asouline, M Wigler
Affiliations

A method for difference cloning: gene amplification following subtractive hybridization

I Wieland et al. Proc Natl Acad Sci U S A. 1990 Apr.

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.

PubMed Disclaimer

References

    1. Cell. 1983 Sep;34(2):557-67 - PubMed
    1. Cell. 1982 Sep;30(2):481-90 - PubMed
    1. Cell. 1984 May;37(1):171-7 - PubMed
    1. Mol Cell Biol. 1984 Sep;4(9):1695-705 - PubMed
    1. Proc Natl Acad Sci U S A. 1985 Jul;82(14):4778-82 - PubMed

Publication types

LinkOut - more resources