Method for rapid amplification of dna

Abstract

The present disclosure relates to methods of DNA amplification with a first primer that has a random sequence of nucleotides at its 3′ end and a generic sequence 5′ of the random nucleotides, as well as a second primer with the generic sequence of the first primer. The disclosure further relates to a method of precipitating DNA on a solid medium to improve DNA amplification. In a preferred embodiment, the presently disclosed methods are used for high-throughput genotyping of DNA samples, such as bloodstains or trace amounts of DNA.

1 - 31 . (canceled) 
     
     
         32 . A method of identifying a polymorphism, the method comprising:
 (a) providing a single reaction mixture comprising:
 (i) a DNA sample; 
 (ii) a first primer comprising a random sequence of nucleotides at its 3′ end and a generic sequence 5′ of the random nucleotides; and 
 (iii) a second primer comprising the generic sequence of the first primer and lacking the random sequence of the first primer; 
   (b) subjecting the DNA sample to DNA amplification by a first DNA polymerase wherein the first primer anneals to the DNA to allow the first DNA polymerase to produce a DNA product;   (c) subjecting the DNA product of step (b) to DNA amplification by heat-stable DNA polymerase wherein the second primer anneals to the DNA product to allow the heat-stable DNA polymerase to produce amplified DNA products; and   (d) analyzing the amplified DNA products of step (c) to identify a polymorphism.   
     
     
         33 . The method of  claim 32 , wherein the polymorphism is a single nucleotide polymorphism (SNP). 
     
     
         34 . The method of  claim 33 , wherein the SNP is identified by Oligonucleotide Ligation Assay (OLA), Doublecode OLA, sequencing, Single Base Extension Assay, allele specific primer extension, or mismatch hybridization. 
     
     
         35 - 46 . (canceled)