Competitive PCR for quantitation of DNA

Details Competitive PCR for quantitation of DNA Competitive PCR for quantitation of DNA

1995-06-02

1998-01-27

Jones, W. Gary

Chemistry: molecular biology and microbiology

Micro-organism, tissue cell culture or enzyme using process...

Preparing compound containing saccharide radical

435810, C12P 1934

Patent

active

057121257

ABSTRACT:
The invention provides a method of determining the amount of target DNA in a sample which comprises the steps of (a) dividing the sample into a plurality of aliquots; (b) adding to each aliquot a known amount of competitor DNA which is the same as the target DNA except that it comprises a recognition site capable of being detected directly or indirectly by a labelled species whereby the ratio of target DNA to competitor DNA is different in the aliquots; (c) co-amplifying the target DNA and competitor DNA in each aliquot by the polymerase chain reaction (PCR) using at least one primer which permits immobilization of the co-amplified DNA, the annealing sites of any PCR primers used being outward of the recognition site; (d) labelling the amplified competitor DNA; (e) before or after step (d), immobilizing the co-amplified DNA where not already immobilized, via at least one PCR primer; (f) assessing the amount of label on the immobilized amplified competitor DNA in each aliquot to provide an indication of the amount of target DNA in the sample.

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