Harvested cells can be stored in -80 degree Celsius for less than a week before RNA isolation or right after harvest.
Add 400 µl Trizol to lyse the cell and incubate at Room Temperature for 5, minutes.
Add 80 µl chloroform (0.2ml per 1ml Trizol)
Vortex the mixture for 15 secs and incubate at Room Temperature for 10 minutes
Centrifuge at 12000rpm / 4 degree Celsius / 15 minutes
Transfer the aqueous phase to the new clean tube and add equal volume of isopropanol
Mix well (short vortex)
Incubate at room temperature for 10 minutes.
Centrifuge at 12000 rpm / 10 minutes/ 4 degree Celsius
Remove isopropanol using pipette keeping the pellet safe
Add 400 µl of 75% ethanol and centrifuge at 7500 rpm/ 5 minutes/ 4 degree Celsius
Pipette out the ethanol keeping the pellet safe
Air dry the pellets for 5-10 minutes
Resuspend the pellet with 20 µl Nuclease Free water.
Measure the concentration of RNA isolated. (SPECTROPHOTOMETRIC ANALYSIS)
Binod G C
I'm Binod G C (MSc), a PhD candidate in cell and molecular biology who works as a biology educator and enjoys scientific blogging. My proclivity for blogging is intended to make notes and study materials more accessible to students.
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