Fast pcr for str genotyping
AssigneeLife Technologies Corporation
InventorsDennis WANG, Lori Hennessy
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Abstract
Disclosed is a method of amplifying a nucleic acid sequence, wherein the method comprises subjecting a reaction mixture to at least one amplification cycle, wherein the reaction mixture comprises a double-stranded nucleic acid and at least two primers capable of annealing to complementary strands of the double-stranded nucleic acid and amplifying at least one short tandem repeat (STR) using a Family A DNA polymerase in a Fast PCR protocol having a two-step amplification cycle in 25 seconds or less. Also disclosed are real-time PCR methods using the two-step protocol and kits for STR profiling using the Fast PCR protocol.
Claims28
A method of amplifying a nucleic acid sequence, wherein the method comprises:
- subjecting a reaction mixture to at least one amplification cycle, wherein the reaction mixture comprises a double-stranded nucleic acid, at least two primers capable of annealing to complementary strands of the double-stranded nucleic acid and amplifying at least one short tandem repeat (STR), and a Family A DNA polymerase, and wherein the at least one amplification cycle comprises:
- denaturing the double-stranded nucleic acid in the reaction mixture;
- and
- annealing the at least two primers to complementary strands of the denatured double-stranded nucleic acid and extending the at least two primers;
- and
- wherein the time to complete one amplification cycle is 25 seconds or less.
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A kit for detection of an STR profile comprising:
- at least two primers wherein said primers are capable of amplifying at least one short tandem repeat (STR) in at least 25 seconds per amplification cycle.
Dependencies are inferred from the claim text.