Download HLA-DRB BigDye™ Terminator Sequencing

Introduction
Overview IMPORTANT
This kit should be used in accordance with guidelines
established by the Committee for Quality Assurance and Standards of the
American Society for Histocompatibility and Immunogenetics.1
The genetic complexity of the DRB region of the human major
histocompatibility complex (MHC) has required development of
molecular typing techniques with increasing levels of resolution.
Methods such as Polymerase Chain Reaction–Sequence-Specific
Oligonucleotide Probe (PCR-SSOP) or Sequence-Specific
Primer–PCR (SSP-PCR) are not always able to discriminate among the
approximately 200 recognized alleles at DRB1.
To address this need, Applied Biosystems has developed a system that
combines a low-resolution SSP-PCR followed by high-resolution allele
typing using automated DNA sequencing.
SSP-PCR The low resolution SSP-PCRs are based on allele group-specific motifs
Amplifications in the first hypervariable region of exon 2. The following allele groups
and genes are amplified with the eleven specific PCR mixes provided:
♦
DR1
♦
DR7
♦
DRB3
♦
DR2
♦
♦
DR3/11/6
♦
DR8/12
♦
DRB4
DR9
♦
DRB5
♦
DR4
♦
DR10
In addition to the group-specific amplification mixes, a control mix is
also provided that amplifies all DRB alleles and genes (DRB*ALL). This
mix can be used as either a positive or negative amplification control.
These PCRs have been optimized for use with AmpliTaq Gold® DNA
Polymerase to ensure specific and efficient reactions.
1.
1996 ASHI Standards for Histocompatibility Testing. American Society for
Histocompatibility and Immunogenetics, 1996 Membership Directory, pp. 17–28.
1