DNA amplification and subtraction techniques

Details DNA amplification and subtraction techniques DNA amplification and subtraction techniques

1988-06-17

2000-08-22

Zitomer, Stephanie W.

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

Involving nucleic acid

435 911, 435 912, 536 242, 536 243, 536 2433, 536 254, C12Q 168, C12P 1934, C07H 2102, C07H 2104

Patent

active

061070238

ABSTRACT:
A method of isolating genomic or RNA-derived duplex fragments which are unique to one of two fragment mixtures. The fragments in positive-source and negative-source mixtures are separately equipped with end linkers, and each mixture is amplified by successive primed-strand replications, using a single primer which is homologous to the associated linker. The second-source linker is biotinylated, and the fragments in this mixture are hybridized in molar excess with the fragments in the positive-source mixture. DNA species which are not hybridized with the biotinylated species, i.e., species that are unique to the positive-source mixture, are isolated after removal of hybridized species by affinity chromatography. Also disclosed is a method of amplifying a mixture of DNA fragments by repeated linker/primer replication.

REFERENCES:
patent: 4683195 (1987-07-01), Mullis et al.
Maniatis et al., Molecular Cloning: A Laboratory Manual, Cold Spring Harbor, NY, 1982.

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