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HLA-DRB sequencing-based typing: an improved protocol which shows complete DRB exon 2 sequences and includes exon 3
A van Dijk1, R Melchers, Y Hilkes
1Department of Pathology, University Medical Centre Utrecht, Utrecht, The Netherlands.
Abstract:
Several human leukocyte antigen (HLA)-DRB protocols for sequencing-based typing have been described. In general, the DRB1 amplification is performed using group-specific amplification primers (GSAPs) located in HVR I or intron 1. Only some protocols include amplification of DRB3, DRB4, and DRB5. However, prior knowledge obtained by alternative methods such as PCR-SSP is preferred for some protocols and a large amount of DNA is often required for adequate typing of HLA-DRB1. We describe a new protocol that uses GSAPs located in the introns, includes also sequencing of exon 3 to enable high resolution. This protocol allows DRB typing without prior knowledge of the HLA type.
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