Method for detecting a nucleic acid base sequence
AssigneeZeneca Limited
InventorsDavid Mark Whitcombe, Jannine Brownie, Stephen Little
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AssigneeZeneca Limited
InventorsDavid Mark Whitcombe, Jannine Brownie, Stephen Little
Estimating the term…
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.
A method for the detection of a diagnostic base sequence in nucleic acid in a sample, which method comprises contacting the sample under hybridizing conditions and in the presence of appropriate nucleoside triphosphates and an agent for polymerization thereof, with a diagnostic primer for the diagnostic base sequence, the diagnostic primer having a non-complementary tail sequence comprising a tag region and a detector region, such that an extension product of the diagnostic primer is synthesised when the corresponding diagnostic base sequence is present in the sample, no extension product being synthesised when the corresponding diagnostic base sequence is not present in the sample and any extension product of the diagnostic primer acts as a template for extension of a further primer which hybridizes to a locus at a distance from the diagnostic base sequence, and contacting the sample with a tag primer which selectively hybridizes to the complement of the tag region in an extension product of the further primer and is extended, and detecting the presence or absence of the diagnostic base sequence by reference to the detector region in the further primer extension product.
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