Dna amplification and subtraction techniques

Details Dna amplification and subtraction techniques Dna amplification and subtraction techniques

C - Chemistry, Metallurgy

12

Q

195/1.13, 150/8.

C12Q 1/68 (2006.01) C12N 15/10 (2006.01)

Patent

CA 1335070

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.

603091

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