Kit for guiding folic acid taking dose for pregnant women and using method of kit
The technology of a kit and detection kit is applied in the biological field to achieve the effect of reducing spontaneous abortion
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[0033] (1) First collect 2ml of peripheral blood from pregnant women, anticoagulate with EDTA, and use SnoMag TM Blood DNA Extraction Kit extracts genomic DNA, and operates in strict accordance with the kit instructions.
[0034] (2) The genotypes of MTHFR C677T, A1298C and MTRR A66G were detected by HRM method. Direct application of LightCycler 480Real-Time PCR System for PCR amplification and HRM analysis. The total PCR reaction system is 10ul, including: genomic DNA 1ul, SsoFast EvaGreen Supermix 5ul, primer (F / R 10uMol) 0.5ul, ddH 2 O added to 10ul. The PCR reaction conditions were: 95°C for 10 min, 95°C for 30S, 60°C for 30S, 72°C for 20S, 45 cycles. HRM analysis conditions: 95°C for 1min, 40°C for 1min, 60°C for 1min, and then the melting curve data was collected from 60°C to 90°C at a rate of 0.1°C / s. The primer sequences are as follows:
[0035] MTHFR C667T:
[0036] F: 5'-GAAGCAGGGAGCTTTGAGGCTGACCTG-3'
[0037] R: 5'-TGCCTTCACAAAGCGGAAGAATGTGTCAGC-3'
[0038]...
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