162
edits
Changes
→Protocol
#Add 1 mL TRIzol reagent to each 2 mL tube.
#Cut ~50-100 mg of tissue. If tissue is in RNAlater, cut at room temperature. If tissue is frozen cut on dry ice. Weigh into a fresh 2mL eppendorf tube and keep on ice.
#Using tissue grinder, homogenize tissue for 3 minutes min at 25Hz (make sure there are no remaining clumps).
#Incubate 5 minutes at room temperature.
#Add 200 uL Chloroform and shake vigourously by hand for 15s. Do not vortex.
#Spin 1 min on max to dry the cartridge. Discard the collection tube and place into a clean recovery tube. Add 100 uL RNAase free water to the center of each tube. This can be adjusted to between 30 and 300 uL elution buffer if necessary.
#Incubate at room temperature for 1min.
#Spin 2 min at 12500 rpm to get purified RNA.
#Quantify the RNA using the nanodrop.
