This kit is used for high-resolution genotyping of HLA-A, HLA-B, HLA-C, HLA-DRB1, and HLA-DQB1 gene loci of the human leukocyte antigen (HLA) coding gene HLA gene complex. The user of this reagent kit is a professional who has received medical laboratory training and familiar with medical molecular biology detection methods and high-throughput sequencing operations. High throughput sequencing, also known as Next Generation Sequencing (NGS), is relative to traditional Sanger Sequencing.
This kit uses the Next Generation Sequencing (NGS) sequencing method to classify HLA genes, and the core of the technology is the linkage reaction sequencing of four fluorescent labeled oligonucleotides. Before sequencing, an amplification mixture was prepared for HLA site specificity, and DNA amplification was performed with Taq polymerase and sample genomic DNA for each site specificity. Mix the amplified products proportionally, break them by enzymatic method, purify them with magnetic beads, and add connectors at the 3 'and 5' ends respectively. These connectors will randomly bind DNA fragments to the glass surface (Flow cell) and undergo bridge amplification on the solid phase surface. This forms thousands of identical single molecule clusters that are used as sequencing templates. Sequencing adopts a method of synthesizing while sequencing. The ddNTP raw materials paired with the template are added, while the unpaired ddNTP raw materials are washed away. The imaging system can capture the fluorescent labeled nucleotides. With the removal of the blocking agent at the 3 'end of DNA, the next round of extension can proceed. The sequencing results were assisted by computer software to complete HLA allele typing.
