Methods for rapid forensic DNA analysis

摘要

The present invention provides methods and primer pairs for rapid, high-resolution forensic analysis of DNA and STR-typing by using amplification and mass spectrometry, determining the molecular masses and calculating base compositions of amplification products and comparing the molecular masses with the molecular masses of theoretical amplicons indexed in a database.

主权项

1. A method for STR typing comprising:
a) amplifying a nucleic acid from a sample with an oligonucleotide primer pair comprising a forward and a reverse primer, each between 13 and 40 nucleobases in length, wherein said forward primer is configured to hybridize within a first conserved region of said nucleic acid and said reverse primer is configured to hybridize within a second conserved region of said nucleic acid, wherein said first and said second conserved regions flank a variable nucleic acid region comprising a STR locus, wherein said amplifying generates at least one amplification product that is between about 45 and about 200 nucleotides in length; b) determining the molecular mass of at least one strand of said at least one amplification product by mass spectrometry; and c) comparing said molecular mass to a molecular mass database comprising a plurality of molecular masses of a plurality of STR-identifying amplification products indexed to said oligonucleotide primer pair and to a reference allele corresponding to said STR locus, wherein a match between said determined molecular mass and a molecular mass comprised in said molecular mass database identifies an STR allele in said sample wherein if no match is identified further comprising: d) calculating the base composition of said at least one strand of said at least one amplification product using said determined molecular mass, wherein said calculated base composition identifies a previously unknown allele of said STR locus; and e) indexing said calculated base composition to said oligonucleotide primer pair, said previously unknown allele, said determined molecular mass and said sample in a database.