Fast PCR for STR Genotyping

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.

We claim: 
     
         1 - 43 . (canceled) 
     
     
         44 . A composition comprising a Pol A enzyme and an amplified short tandem repeat (STR) locus, wherein the enzymatic activity of the Pol A enzyme present is such that a doubling of enzymatic activity increases the presence of an amplified STR locus with a split peak. 
     
     
         45 . The composition of  claim 44 , wherein the Pol A enzyme is derived from  Thermus aquaticus.    
     
     
         46 . The composition of  claim 45 , wherein the enzymatic activity is 7.5 Units. 
     
     
         47 . The composition of  claim 45 , wherein the enzymatic activity is from 7.5 Units to 10 Units. 
     
     
         48 . The composition of  claim 44 , wherein the amplified STR locus is D5S818. 
     
     
         49 . The composition of  claim 44 , wherein the amplified STR locus is D8S1179.