GeneQuery™ HLA-A 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-A PCR+Sanger SBT Typing Kit (SBT-A) 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-A kit contains a ScienCell proprietary HLA-A amplification primer set (A_ampF primer mix and A_ampR primer mix; Cat #GK891a) that can amplify the complete HLA gene sequence from the enhancer-promoter region to the 3' UTR region, and six sequencing primer mixes: A_2F (Cat # GK891b), A_2R (Cat # GK891c), A_3F (Cat # GK891d), A_3R (Cat # GK891e), A_4F (Cat # GK891f), and A_4R (Cat # GK891g) 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 #GK891r). The GDR can be used to monitor the quality of the reagents and the instruments. The HLA-A alleles in GK891r are HLA-A*02:01 and HLAA*25:01.

GeneQuery™ HLA-A PCR+Sanger SBT Typing Kit (SBT-A) 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:

• #GK891a - HLA-A amplification primer set (A_ampF and A_ampR) -400 uL - Storage: -20°C
• #GK891b - A_2F sequencing primer mix, 10 uM - 200 uL - Storage: -20°C
• #GK891c - A_2R sequencing primer mix, 10 uM - 200 uL - Storage: -20°C
• #GK891d - A_3F sequencing primer mix, 10 uM - 200 uL - Storage: -20°C
• #GK891e - A_3R sequencing primer mix, 10 uM - 200 uL - Storage: -20°C
• #GK891f - A_4F sequencing primer mix, 10 uM - 200 uL - Storage: -20°C
• #GK891g - A_4R sequencing primer mix, 10 uM - 200 uL - Storage: -20°C
• #GK891h - Nuclease-free H2O - 4 mL - Storage: 4°C
• #GK891r - Human genomic DNA reference (GDR), ~50 ng/uL (HLA-A*02:01 and HLA-A*25:01) - 10 uL - Storage: -20°C
• #MB6038-1 - 2X EmeraldNStart HiFi Marathon PCR Master Mix, 1 mL - 2 vials - Storage: -20°C