Specific primer group, test kit and test method for testing polymorphic sites of human MTHFR and MTRR genes
A gene polymorphism, human detection technology, applied in biochemical equipment and methods, recombinant DNA technology, microbial determination/inspection, etc. It can reduce the pain, use less blood, and reduce economic expenses.
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Embodiment 1
[0039] Embodiment 1: the preparation of kit
[0040] 1. Design and synthesis of primers and probes
[0041] For the MTHFR (C677T), MTRR (A66G), and MTHFR (A1298C) gene loci in the human genome (for the sequence, refer to the human genome sequence published in the NCBI database), specific primers and probes were designed using Primer Premier 3.0 software. Specific primers and probe sequences are shown in Table 1:
[0042] Table 1:
[0043]
[0044] 2. Selection of reference substance
[0045]The wild-type control is the wild-type plasmid mixture of the three gene loci of MTHFR (C677T), MTHFR (A1298C), and MTRR (A66G); the negative control is 1*TE buffer or nuclease-free water.
[0046] 3. Composition of PCR reaction solution
[0047] The PCR reaction solution includes the above-mentioned specific primers and probes, wherein:
[0048] The lyophilized powder in the reaction tube contains M677 forward primer F1, M677 reverse primer R1, M677 probe P1, M1298 forward primer F...
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