Preparation of RNA Samples from Mouse Tissues: Difference between revisions
updated for purelink kits. |
added ethanol to materials and prepared TRIzol tubes first |
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*Chloroform (in solvent cabinet) | *Chloroform (in solvent cabinet) | ||
*Label tubes, for each sample need 3 eppies, at least one of which is 2 mL and one of which is a recovery tube from the PureLink kit. | *Label tubes, for each sample need 3 eppies, at least one of which is 2 mL and one of which is a recovery tube from the PureLink kit. | ||
*70% Ethanol make with RNAase free water and 100% Ethanol. | |||
==Protocol== | ==Protocol== | ||
#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 | #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 ~30s until homogeneous (no clumps). | #Using tissue grinder, homogenize tissue for ~30s until homogeneous (no clumps). | ||
#Incubate 5 minutes at room temperature. | #Incubate 5 minutes at room temperature. | ||