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MicroRNA and TotalRNA Purification from Liver Tissue

Instrument: Precellys® Evolution with 7mL adapter

Sample: rat liver

Protocol: 

  • Precellys kits: CK28_7mL (KT03961-1-302.7)
  • Rat liver (50-100 gm)
  • Buffer: 1 mL of RNAdvance Tissue Lysis buffer and anti foaming DX Reagent
  • Adjust lysate to 10 mg/400μL of buffer and then digest with proteinase K at 37°C for 25 minutes, followed by RNA extraction using the RNAdvance Tissue miRNA protocol
  • Run the Precellys at 6500 rpm, one cycle of 20 seconds, and 600 rpm for a 2nd cycle of 20 seconds. 
  • RNA concentration was measured with a NanoDrop 2000 spectrophotometer
  • Let-7c miRNA gene expression was measured using the Taqman microRNA assay

Results: 

Liver tissue replicates were used to evaluate miRNA extraction efficiency. The average yield was between 25-30 μg per 5 mg liver tissue. The average cycle threshold (Ct), value for let-7c miRNA expression in RNA samples 25 cycles. 

The average RNA yield prepared from the Precellys homogenizer.

Overlay of let-7c gene expression from the RNAdvance Tissue Kit.