MicroRNA and TotalRNA Purification from Liver Tissue
Instrument: Precellys® Evolution with 7mL adapter
Sample: rat liver
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
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.