Long-range PCR amplification as an alternative strategy for characterizing novel HLA-B alleles
Curran, M.D., Williams, F., Earle, J.A.P., Rima, B.K., Dam, M.G., Bunce, M. and Middleton, D. (1996) Long-range PCR amplification as an alternative strategy for characterizing novel HLA-B alleles. European Journal of Immunogenetics, 23 (4). pp. 297-309.
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We have developed a simple, rapid and reliable method for specifically amplifying and cloning full-length HLA-B genes from genomic DNA. Using this methodology we characterized three alleles of interest at the molecular level. Two of the alleles appeared in our routine class I PCR-SSOP typing system, a variant of B*5801 found in the Daudi cell line and RCE 56 and a variant of B*4101 found in a number of volunteer donors on our Bone Marrow Donor Registry. The third, a variant B35 allele found in RCE 80, was first identified as unusual by serology. Our sequencing analysis of exon 2 and exon 3 identified two of these alleles as the recently reported novel HLA-B*5802 and HLA-B*4102 alleles, while the third represents a new B35 allele officially designated B*3513.
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