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Improving RNA Extraction from Mouse Tissues using the Precellys24 Dual

Instrument: Precellys® 24 Dual

Sample:  Heart, liver, spleen

Protocol:

  • Precellys kits: CK14 2mL (KT03961-1-003.2) for heart, liver and CKMix 2mL (KT03961-1-009.2) for spleen
  • Samples: Heart, liver, spleen (10-30mg)
  • Buffer: 1mL of Trizol reagent
  • Run the Precellys24-Dual at 5000 rpm, 1x30 sec
  • Pulverization: Mortar and pestle, followed by homogenization using a needle and syringe in Trizol.
  • Methods: Mouse tissues were removed and snap frozen in liquid nitrogen or dropped into RNAlater prior to homogenization.

Results:

RNA integrity and yield was evaluated using an Agilent Bioanalyzer, and an average RIN (RNA integrity number) value of 9 (highest score is 10) was observed using the Precellys, with RNA concentration falling between 400-1800 ng/ul. The gel displays clean 18S and 28S rRNA bands with little or no degradation (Figure 1A). RIN values of “N/A” were observed after pulverization, which is indicative of highly degraded RNA as observed on the gel (Figure 1B).

The Precellys24-Dual significantly improves RNA integrity and yield compared to our previous pulverization method. In addition, the time spent on sample homogenization was decreased from one hour to 30 seconds

 
RNA integrity after sample homogenization on the Precellys24-Dual (A) or using pulverization (B).