Beta-thalassemia gene amplification kit and method based on recombinase polymerase amplification technology
A recombinase polymerase, thalassemia technology, applied in biochemical equipment and methods, microbial determination/inspection, etc., can solve the problems of PCR amplification dependence and slow amplification rate, and achieve high sensitivity and fast amplification time. Effect
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[0055] (1) Primer design and optimization
[0056] Using bioinformatics knowledge and related bioinformatics software, analyze the β-globin gene sequence, design specific RPA amplification primers for RPA amplification, and analyze their sequences, especially the primer pair After the sequence analysis of the combination, the qualified primers were selected for experimental verification and optimization. Alternative primers are listed in Table 4.
[0057] Table 4 alternative PCR primer sequences (both 5'-3')
[0058]
[0059]
[0060] F in the table indicates the upstream primer, R indicates the downstream primer; bio indicates the modification of Biotin at the 5' end.
[0061] (1) Prepare each primer to 10 μM and set aside. Using human genomic DNA (extracted from peripheral blood, negative for thalassemia, Guangdong) as a template, the primers are optimized according to the following process: the present invention divides the β-globin gene into three fragments for am...
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