RNA isolation from cells | RNA quantification
RNA isolation from cells Protocol: Aspirate off media, wash once with ice cold PBS (1-2) mL. Aspirate off PBS, add 1 mL Trizol . Scrape the plate with a cell scraper, collect the cell lysate to 1.5 mL E-tube. Incubate RT for 5mins. Centrifugation 4°C, 13000 rpm for 10 mins. Aliquot supernatant to new E-tube. Add 200 µL chloroform (200 µL + 800 µL=1 mL), vortexing. Incubate RT for 10-15 mins (vortex every 2 mins) Centrifugation 4°C, 13000 rpm for 15 mins. Separate aqueous phase and transfer to new E-tube. Add isopropanol (equal volume of aqueous phase) Inverting, incubate RT for 5-10 mins. Centrifugation 4°C, 13000 rpm for 10 mins. Remove supernatant and add 700 µL 70%EtOH to wash the pellet. Centrifugation 4°C, 13000 rpm for 5 mins. Remove supernatant and add 1 mL 70%EtOH to wash the pellet. Centrifugation 4°C, 7000 rpm for 10 mins. Remove supernatant, spin down. Use yellow tip remove supernatant completely. Air dry rapping with poly for 5-10 mins. Add DE