Subtractive hybridization and capture methods and kits for diffe

Details Subtractive hybridization and capture methods and kits for diffe Subtractive hybridization and capture methods and kits for diffe

1997-01-09

1998-03-10

Myers, Carla J.

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

Involving nucleic acid

435 912, 536 242, 536 2433, 935 8, 935 78, C12Q 168, C12P 1934, C07H 2104

Patent

active

057260222

ABSTRACT:
The invention disclosed here allows for the differential isolation of nucleic acid sequences that are present in one nucleic acid population and not in another. The method is based upon using restriction endonucleases to digest two populations of nucleic acid, preferably cDNAs, preferably ligating different sets of adaptors to each of the two nucleic acid populations, followed by hybridization, restriction digestion and isolation of the desired molecules. The unique aspects of this invention include the use of a restriction enzyme to isolate the target duplex DNA molecule from a hybridization mixture. Certain embodiments of the invention include the direct or indirect incorporation of a capture molecule or ligand (e.g., biotin, dioxigenin) within the amplified nucleic acid fragments, which allows for a system in which molecules can be rescued from both the captured population as well as the effluent or otherwise uncaptured population.

REFERENCES:
patent: 5436142 (1995-07-01), Wigler et al.
patent: 5501964 (1996-03-01), Wigler et al.
Lisitsyn et al., Science 259:946 (1993).

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