Zeneca Limited

Syngenta Corporation

Stephen Little

David Mark Whitcombe

Jannine Brownie

Alec John Jeffreys

Richard Mark Ferrie

Neil James Gibson

Kemal Haque

Jane Theaker

Jayne C Fox

A method for detecting allelic imbalance (AI) in sample nucleic acid, which method comprises providing multiple copies of a target nucleic acid region present in the sample, the target nucleic acid region comprising a marker of heterozygosity, separating the multiple copies into individual strands then allowing the individual strands to reanneal under conditions which permit the formation of homoduplexes and heteroduplexes, removing any heteroduplexes so formed, subjecting the remaining homoduplexes to the above steps of separation, reannealing and heteroduplex removal one or more times so that any difference in the initial ratio of allelic variants is amplified, and detecting the presence or absence of AI by reference to any difference in allele ratio so detected.

Method for detecting allelic imbalance

A method for the detection of a target nucleic acid, which method comprises contacting template nucleic acid from a sample with (i) a signalling system and (ii) a tailed nucleic acid primer having a template binding region and the tail comprising a linker and a target binding region, in the presence of appropriate nucleoside triphosphates and an agent for polymerization thereof, under conditions such that the template binding region of the primer will hybridize to a complementary sequence in the template nucleic acid and be extended to form a primer extension product, separating any such product from the template whereupon the target binding region in the tail of the primer will hybridize to a sequence in the primer extension product corresponding to the target nucleic acid, and wherein any such target specific hybridization causes a detectable change in the signalling system, such that the presence or absence of the target nucleic acid in the sample is detected by reference to the presence or absence of a detectable change in the signalling system.

Methods for detecting target nucleic acid sequences

A method for the detection of diagnostic base sequences in a sample nucleic acid comprises contacting the sample in the presence of appropriate nucleoside triphosphates and an agent for polymerization thereof with a diagnostic primer for the diagnostic base sequence, the primer having a tail sequence comprising a tag region and a detector region such that an extension product of the primer is synthesized when the corresponding base sequence is present in the sample, and any extension product of the diagnostic primer acting as a template for extension of a further primer which hybridizes to a locus at a distance from the diagnostic base sequence. The sample is contacted with a tag primer which selectively hybridizes to the complement of the tag sequence in an extension product of the further primer and is extended, and the presence or absence of the diagnostic base sequence is detected by reference to the detector region in the further primer extension product.

Method for detecting a nucleic acid base sequence

The present invention relates generally to a method of characterizing a sample of genomic DNA and to nucleotide sequences employed in the method as well as kits comprising these. In particular the invention involves the use of primers which selectively prime specific type(s) of internal repeat unit in a tandemly repeated region. The method of the invention is particularly useful in forensic or paternity studies and provides individual sample codes suitable for computerized storage on, and retrieval from, a database.

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Northwich, United Kingdom

Stephen Little