A primer, probe and kit for detecting human mthfr and mtrr gene mutations
A technology for detecting people and kits, which is applied in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc. , can not fully cover the problem of gene mutation sites, etc., to achieve the effect of intuitive typing, improving analysis performance, and accurate typing
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Embodiment 1
[0067] Example 1 Production and assembly of a kit for detecting human MTHFR and MTRR gene mutations:
[0068] Primers and probes are:
[0069] MTHFR C677T Primer 1 (SEQ ID NO: 1);
[0070] MTHFR C677T Primer 2 (SEQ ID NO:2);
[0071] MTHFR C677T probe 1 (the nucleotide sequence is SEQ ID NO: 7), ROX-TGTCTGCGGGAGCCGATTTCATCATC-BHQ2;
[0072] MTHFR A1298C Primer 3 (SEQ ID NO:3);
[0073] MTHFR A1298C Primer 4 (SEQ ID NO:4);
[0074] MTHFR A1298C probe 2 (the nucleotide sequence is SEQ ID NO: 8), FAM-CCAGTGAAGAAAGTGTCTTT-MGB;
[0075] MTRR A66G Primer 5 (SEQ ID NO:5);
[0076] MTRR A66G Primer 6 (SEQ ID NO:6);
[0077] MTRR A66G probe 3 (nucleotide sequence is SEQ ID NO: 9), VIC-CAGAAGAAATGTGTGAGCAAG-MGB.
[0078] Kit includes:
[0079] Reaction solution (see Table 2 for detailed composition), enzyme (see Table 3 for detailed composition), positive quality control product (see Table 4 for detailed composition) and blank quality control product (see Table 4 for detailed co...
Embodiment 2
[0090] Embodiment 2 The kit provided by the present invention is used for the detection of clinical samples
[0091] 30 cases of clinical samples were tested using the 48-person kit in Example 1 of the present invention. The composition of the kit is shown in Table 5.
[0092] table 5
[0093] kit components Component Specifications Number of components The reaction solution 1109μL / tube 1 tube enzyme 53μL / tube 1 tube positive control 15μL / tube 1 tube blank quality control 1.5mL / tube 1 tube
[0094] 1. Nucleic acid extraction
[0095] Take 200 μL whole blood samples and 200 μL blank quality control products, use the magnetic bead method blood genome extraction kit (Tiangen Biochemical Technology (Beijing) Co., Ltd., DP329) in the automatic nucleic acid extraction instrument (Tiangen Biochemical Technology (Beijing) Co., Ltd., TGuideS32) for nucleic acid extraction, the extraction is carried out according to the instruction...
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