Primer group, application, product and method for detecting SNP (single nucleotide polymorphism) loci related to drug metabolism capacity of rheumatic immunological diseases
A technology of metabolic capacity and rheumatic immunity, which is applied in the fields of biochemical equipment and methods, recombinant DNA technology, and microbial determination/inspection, etc., can solve problems such as lack of throughput, and achieve lower detection costs, higher accuracy, and shorter detection cycles. Effect
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Embodiment 1
[0100] Example 1: Design and optimization of primers, and establishment of reaction system
[0101] Through the primer design software (Assay Design Suite) of the MassARRAY website, adjust the relevant parameters, complete the PCR and UEP primer design of 44 loci (including a repeat detection locus rs2231142 and sex identification locus AMEL), and export the designed primers And each parameter file, and synthesize primers. Prepare amplification primer MIX and extension primer MIX according to the primer configuration table, and fine-tune the extension primer MIX until it meets the requirements. Primer testing and optimization were then performed. Specific steps are as follows:
[0102] (1) Dilute the genomic DNA sample to 10ng / μL, and prepare the PCR reaction MIX according to the following table, where the PCR PrimerMix is a mixture of two sets of amplification primers with different sites (the following is the amount of a single sample):
[0103] Table 3 PCR reaction sys...
Embodiment 2
[0151] Verification of embodiment 2 reaction system
[0152] In the process of system optimization, 2 to 3 samples of each detection site were verified by Sanger sequencing, and the comparison results were consistent, confirming that the detection results were accurate. After confirming the optimal reaction system, the following experiments were carried out:
[0153] (1) Accuracy experimental verification plan: select one sample from each of the 43 sites for Sanger sequencing, and compare the results of Sanger sequencing and MassARRAY. If the consistency is greater than 95%, the verification is passed;
[0154] (2) Precision experiment verification plan: pick 2 peripheral blood samples and 2 oral swab samples, repeat 3 times for each sample to conduct a batch of detection, a total of 3 batches were detected, the precision between batches and batches If the consistency of internal precision is greater than 95%, the verification is passed;
[0155] (3) Experimental operator co...
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