Isolation of RNA from whole tissue

15. Remove the tissues from the -80°C freezer and place on dry ice.

16. Pre-chill the stainless steel mortar and pestle sets on dry ice. Place the frozen tissue into the cold mortar. Place the pestle onto the mortar and pulverize the tissue into a frozen powder by forcefully striking with a 2-pound hammer.

17. Transfer the frozen, pulverized tissue as quickly as possible into a 1.5 ml microcentrifuge tube that has been pre-chilled on dry ice. Do not let the pulverized tissue thaw. A sterile disposable surgical blade is ideal for transferring the powdered tissue. Use one new blade per tissue sample.

18. Place the tubes containing the pulverized tissue on dry ice until all of the samples are ready for lysis.

19. Proceed with steps 4-14 to isolate the RNA.

Use the mortar and pestle sets only once per tissue. Clean the mortar and pestle sets with soap and water, dry and re-chill on dry ice prior to isolating any additional tissues.

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