Patent ID: 6287764
Filing Date: 2001-09-11
Classification: C12Q

Abstract:
A method for typing HLA class I alleles, comprising the steps of:amplifying HLA-A, -B, or -C alleles so as to provide a primary amplicon, wherein the primary amplicon comprises amplified exons 2 and 3 of the HLA-A, -B, or -C alleles in a locus specific manner;producing at least two secondary HLA-A, -B, or -C locus specific amplicons corresponding to each of the exons 2 and 3 of the primary amplicon, wherein the at least two secondary amplicons are produced by amplifying the primary amplicon by means of using the primary amplicon as a template and nested polymerase chain reaction primers, thereby independently framing each of exon 2 and exon 3 with nested primers in the neighboring introns;preparing the at least two secondary amplicons for sequencing, wherein the at least two secondary amplicons are provided with an anchoring moiety attached to a first terminus of each of the at least two secondary amplicons and a sequencing primer site attached to a second terminus of each of the at least two secondary amplicons;attaching the anchoring moiety of each of the at least two secondary amplicons to solid supports; andDNA sequencing each of the at least two secondary amplicons.