RNA Precipitation

  1. Treat trizol 300 ul/each well (format 12 well plate)
  2. Incubate 10 min on-ice
  3. Transfer trizol-cell lysate to e-tube [Pre-marked]
  4. Add 30 ul BCP solution into e-tube, vortex, and then keep on ice for 5 min
  5. Spindown at 12.000 g for 15 min
  6. Take e-tube back to ice, transfer Aquous phase (the upper supernatant contain RNA) approximately 120 ul to a new e-tube [Pre-marked]
  7. Put same volumes of Isopropanol with supernatant into e-tube
  8. Invert 5 times gently and then incubate for 10 min at room temperature
  9. Spindown at 12.000 g for 8 min
  10. Look at the visible pellets, pour out whole supernatant
  11. Add 1 mL 75% EtOH in DEPC-treated water into e-tube
  12. After inverting, check pellets whether resuspended or not
  13. Spindown 7500 g for 5 min 14 Remove supernatant using pipet and dry at room temperature
  14. Resuspend the pellet with 30 ul DEPC-treated water

cDNA synthesis

  1. Measure