Subtractive hybridization techniques for identifying differentia

Details Subtractive hybridization techniques for identifying differentia Subtractive hybridization techniques for identifying differentia

1997-07-24

1999-08-10

Ketter, James

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

Involving nucleic acid

435 912, 536 231, 536 242, 536 2433, C12Q 168, C12P 1934, C07H 2104

Patent

active

059357886

ABSTRACT:
The present invention provides gene subtraction methods that can be used to identify and isolate from hybridization mixtures differentially expressed nucleic acid and/or commonly expressed nucleic acid between two populations of nucleic acid. The methods of the present invention employ a restriction endonuclease recognition site that is also recognized in whole or in part by a second restriction endonuclease (e.g., HinPI and BssHII or HhaI and HinP1I) to create different ends (e.g., sticky ends) between the first and second nucleic acid populations. In addition, the methods of the present invention use a selective ligation step, a biotinylated nucleotide extension step, or both to differentiate between homoduplexes, which allow for the identification of sequences that are different between the two nucleic acid populations, and heteroduplexes, which allow for the identification of sequences that are common to the two nucleic acid populations.

REFERENCES:
patent: 5436142 (1995-07-01), Wigler et al.
patent: 5501964 (1996-03-01), Wigler et al.
patent: 5726022 (1998-03-01), Burmer
Yang et al. Cloning differentially expressed genes by linker capture subtraction. Analytical Biochemistry. vol. 237: 109-114. May 21, 1996.
Velculescu et al. Serial analysis of gene expression. Science. vol. 270: 484-487. Oct. 20, 1995.
Lisitsyn, N., et al., "Cloning the Differences Between Two Complex Genomes," Science, vol. 259 (Feb. 12, 1993), pp. 946-951.

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