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-DRB1 PCR+Sanger SBT Typing Kit (SBT-DRB1) 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-DRB1 kit contains a ScienCell proprietary HLA-DRB1 amplification primer set (R_ampF primer mix and R_ampR primer mix; Cat #GK896a) that can amplify a region in the human genome that covers exons 2-3 of HLA-DRB1 gene, and 6 sequencing primer mixes (R_2F, R_2R, R_3F, and R_3R; Cat #s GK896b-e) 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 #GK896r). The GDR can be used to monitor the quality of the reagents and the instruments. The HLA-DRB1 alleles in GK896r are HLA-DRB1*03:01 and HLA-DRB1*15:01.
GeneQuery™ HLA-DRB1 PCR+Sanger SBT Typing Kit (SBT-DRB1) 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:
- #GK896a - HLA-DRB1 amplification primer set (R_ampF and R_ampR) - 400 uL - Storage: -20°C
- #GK896b - R_2F sequencing primer mix, 10 uM - 200 uL - Storage: -20°C
- #GK896c - R_2R sequencing primer mix, 10 uM - 200 uL - Storage: -20°C
- #GK896d - R_3F sequencing primer mix, 10 uM - 200 uL - Storage: -20°C
- #GK896e - R_3R sequencing primer mix, 10 uM - 200 uL - Storage: -20°C
- #GK896h - Nuclease-free H2O - 4 mL - Storage: 4°C
- #GK896r - Human genomic DNA reference (GDR), ~50 ng/uL (HLA-DRB1*03:01 and HLA-DRB1*15:01) - 10 uL - Storage: -20°C
- #MB6038-1 - 2X EmeraldNStart HiFi Marathon PCR Master Mix, 1 mL - 2 vials - Storage: -20°C
