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
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.