Specific miRNAs in peripheral plasma for pre-eclampsia and application thereof
A preeclampsia-specific technique applied in the field of molecular biology
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[0021] Preeclampsia patients (PE, n=22) and non-preeclampsia pregnant women (Control, CTR, n=40) who delivered in the obstetrics department of our hospital during 2019-2020 were selected. Termination of pregnancy by uterine delivery.
[0022] 1. Sample preparation: Anticoagulate 5ml of peripheral blood with EDTA, centrifuge at 1000g for 5min at 4°C, collect plasma and store it in a low-temperature refrigerator for later use;
[0023] 2. RNA extraction: take 300μl plasma, centrifuge at 4°C, 1000g for 5min; take 250μl upper plasma, add TRI successively and chloroform, centrifuge to take 400 μl of the supernatant, add isopropanol and glycogen, discard the supernatant after centrifugation; add 75% ethanol to wash the precipitate and dry it, and finally add 25 μl of RNase-free water preheated to 65°C to dissolve the RNA.
[0024] 3. cDNA synthesis: mix equal amounts of reverse transcription primers into 2μM mixed reverse transcription primers; thaw the reverse transcription kit o...
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