Product Name
RBC NGS Typing Kits
Application
This kit is used for rapid high-resolution genotyping of human red blood cell antigen (RBCA) coding gene loci.
Experimental Principles
This kit uses the Next Generation Sequencing (NGS) sequencing method to type the RBCA gene, and prepares amplification mixture for RBCA site specificity. The mixture is then amplified with Q5 Taq polymerase and sample genomic DNA for each site specific DNA amplification. Mix the amplified products proportionally, break them by enzymatic method, purify them with magnetic beads, and add adaptor to connectors at the 3 'and 5' ends . These adaptors 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 RBCA allele typing.
In 1900, Landsteiner discovered blood groups A, B, and O; In 1902, two students of Landsteniner discovered relatively rare type AB blood; In 1927, the original letter naming of Landstenier was adopted internationally, and the ABO blood group system was officially established. Four types of blood types were determined: A, B, O, and AB; In 1940, Landsteiner et al. discovered the presence of Rh blood group antigens on red blood cells in rhesus macaques and most human bodies, hence the name
Production Of ABO Blood Group System Antigens
It exists on the red blood cell membrane and is also distributed on white blood cells, platelets, and other tissue cells; The fetus can detect ABH antigen at 5-6 weeks and fully develop at 18 months; Around the age of 20, reaching its peak, the antigenicity of the elderly decreases; Blood type antigens generally remain unchanged throughout life. 
ABO Blood Group System Antigen Inheritance
Located in the long arm of chromosome 9 (9q34)
Three alleles: A, B, O; Among them, A and B are dominant genes, and O is invisible genes; Each parent inherits one gene to their offspring, which can form six genotypes and four phenotypes.
ABO Blood Group System Subtypes
It refers to changes caused by certain differences in the structure and performance of antigens or the number of antigenic sites that belong to the same blood group antigen
Mainly including: A1, A2, A1B, A2B
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Inspection Method
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Note
1. General Safety Instructions
1.1 Only for in vitro diagnosis.
1.2 It must be operated by well trained and authorized personnel.
1.3 All reagents and samples should be considered as potential sources of pollution and appropriate preventive measures should be taken.
1.4 Before and after PCR, it should be conducted in different experimental spaces and using different samplers, and cannot be mixed.
1.5 Gum stained ethidium bromide is a potential carcinogen and should not come into direct contact with the skin. Wear protective gloves during the experiment
1.6 During the UV gel imaging, the eyes should not look directly at the UV light source, and should wear anti UV eyes.
1.7 Pay attention to the expiration date of the reagent and do not use expired reagents.
1.8 Do not use reagents that are cloudy or may be contaminated by microorganisms.
1.9 The used PCR microplate should be considered as a potential source of pollution and, like the agarose gel stained with ethidium bromide, should be destroyed in accordance with relevant national regulations.
1.10 The safety data sheet for experimental reagents and consumables can be obtained from Beijing ROSE Genomics Corp.Ltd
2. Experimental Precautions
2.1 Due to the high sensitivity of PCR experiments, correct operation should be carried out to ensure the purity of the reagents. In order to obtain the correct typing results, please strictly follow the operating instructions.