Pharmaceutical application of hydrogen sulfide donor
A hydrogen sulfide donor, sodium hydrosulfide technology, applied in the field of biomedicine, can solve the problems of no relevant research, loss of smooth muscle, etc., and achieve the effect of reducing mortality, reducing the level of inflammatory factors, and reducing the incidence rate
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[0017] 1. RNA extraction and Q-PCR
[0018] Extraction of total RNA
[0019] (1) After the vascular tissue was washed with ice-cold PBS, 1 mL of Trizol was added, the cells were pipetted and transferred to an RNase-free Eppendorf centrifuge tube, and placed on ice for 5 minutes;
[0020] (2) Add 200 μL of chloroform, shake vigorously to mix, and place on ice for 2-3 minutes;
[0021] (3) 12000rcf, 4°C, 15 minutes;
[0022] (4) Transfer 400 μL of the supernatant to a new Eppendorf centrifuge tube, add an equal volume of isopropanol, mix by inverting, and place on ice for 10 minutes;
[0023] (5) 12000rcf, 4°C, 10 minutes, discard the supernatant;
[0024] (6) Add 75% ethanol diluted with DEPC water, 7500rcf, 4°C, 5 minutes;
[0025] (7) Take the supernatant, dry it at room temperature, add 20 μL DEPC water to dissolve, and measure the concentration of RNA with a UV spectrophotometer;
[0026] (8) Store the purified RNA samples at -80°C.
[0027] reverse transcription
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