GeneQuery™ HLA-B PCR+Sanger SBT Typing Kit

The human leukocyte antigen (HLA) complex is a gene complex located on the short arm of human chromosome 6. The HLA complex is the human version of the major histocompatibility complex (MHC) which helps the immune system distinguish host cells from invaders. It is the most polymorphic locus in the human genome, and has been implicated in many pathologies including autoimmunity disorders and cancer. GeneQuery™ HLA-B PCR+Sanger SBT Typing Kit (SBT-B) utilizes the Sequence-Based Typing (SBT) method, which is the gold standard for HLA typing. This method involves PCR amplification of HLA genes, followed by Sanger sequencing of the amplicons. The acquired sequences are aligned to an HLA allele database using the SOAPTyping software (BGI Genomics) to obtain the 4-digit genotypes of the samples. The two haplotypes from both parents can be typed simultaneously using this kit and the SOAPTyping software. SBT-B kit contains a ScienCell proprietary HLA-B amplification primer set (B_ampF primer mix and B_ampR primer mix; Cat #GK892a) that can amplify a region in the human genome that covers exons 2-4 of HLA-B gene, and 6 sequencing primer mixes (B_2F, B_2R, B_3F, B_3R, B_4F, and B_4R; Cat #s GK892b-g) that can be used in the following Sanger sequencing of the PCR product. Please refer to Figure 2 for the binding location of the primers on the human genome. Each GeneQuery™ HLA typing kit contains a human Genomic DNA Reference (GDR) sample with known HLA alleles (Cat #GK892r). The GDR can be used to monitor the quality of the reagents and the instruments. The HLA-B alleles in GK892r are HLA-B*08:01 and HLA-B*18:01.

GeneQuery™ HLA-B PCR+Sanger SBT Typing Kit (SBT-B) utilizes the Sequence-Based Typing (SBT) method, which is the gold standard for HLA typing. This method involves PCR amplification of HLA genes, followed by Sanger sequencing of the amplicons. The acquired sequences are aligned to an HLA allele database using the SOAPTyping software (BGI Genomics) to obtain the 4-digit genotypes of the samples. The two haplotypes from both parents can be typed simultaneously using this kit and the SOAPTyping software.

Kit Components:

• #GK892a - HLA-B amplification primer set (B_ampF and B_ampR) - 400 uL - Storage: -20°C
• #GK892b - B_2F sequencing primer mix, 10 uM - 200 uL - Storage: -20°C
• #GK892c - B_2R sequencing primer mix, 10 uM - 200 uL - Storage: -20°C
• #GK892d - B_3F sequencing primer mix, 10 uM - 200 uL - Storage: -20°C
• #GK892e - B_3R sequencing primer mix, 10 uM - 200 uL - Storage: -20°C
• #GK892f - B_4F sequencing primer mix, 10 uM - 200 uL - Storage: -20°C
• #GK892g - B_4R sequencing primer mix, 10 uM - 200 uL - Storage: -20°C
• #GK892h - Nuclease-free H2O - 4 mL - Storage: 4°C
• #GK892r - Human genomic DNA reference (GDR), ~50 ng/uL (HLA-B*08:01 and HLA-B*18:01) - 10 uL - Storage: -20°C
• #MB6038-1 - 2X EmeraldNStart HiFi Marathon PCR Master Mix, 1 mL - 2 vials - Storage: -20°C