Isolation of Total RNA from Animal Cells use RNeasy Mini Kit
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实验试剂
实验步骤
1) Trypsinize and collect cells, aspirate medium, and wash cells with PBS twice.
2) Determine the number of cells.
1) Thaw frozen cell pellets slightly.
4. Add 1 volume (usually 350 µl or 600 µl) 70% ethanol, mix using pipet. Do not centrifuge.
8. Add the DNase I incubation mix (80 µl) directly onto the RNeasy membrane, 20–30°C for 15 min.
9. Run 700µl RW1 through the mini column, 15 seconds at 12,000 rpm.
11. Spin another 500µl RPE through the column for 2 minutes at maximum speed (13,200 rpm for us).
注意事项
1. All steps performed at room temperature.
2. RNA can be stored at –80°C for short time.
3. At step 8, it is needed to prolong the treated time for cell >5 x 106.