GenCatch™ Total RNA Extraction Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	- If A260/A280 is low Use ddH2O of acidic pH to dilute RNA sample for spectrophotometric analysis Use 10 mM Tris-HCl of pH 7.5 or TE buffer to dilute RNA sample.	- If Little or no RNA eluted ,Reduce the amount of starting sample and perform more disruption and homogenization.

Protocol tips
- If A260/A280 is low Use ddH2O of acidic pH to dilute RNA sample for spectrophotometric analysis Use 10 mM Tris-HCl of pH 7.5 or TE buffer to dilute RNA sample.
Downstream tips
- If Little or no RNA eluted ,Reduce the amount of starting sample and perform more disruption and homogenization.

Publication protocol

Rat (Rattus norvegicus) liver tissue was donated by Clair Eckersell (Brigham Young University - Idaho). Identical to the cell RNA extraction protocols, both kit procedures followed the manufacturer’s instructions using 30 mg of tissue with no modification and the subsequent RNA obtained was stored at −80°C. For the silica column procedure, approximately 30 mg of tissue was placed in 2 ml centrifuge tubes (VWR, Radnor, PA, 89000-010) followed by the addition of 600 µl of Buffer A. Tissue was disrupted using a homogenizer (Omni International, Kennesaw, GA, TH115). The resultant mixture was centrifuged at 8000 G for 3 min and the supernatant was transferred to a new 2 ml centrifuge tube and one volume of 70% ethanol was added. The mixture was then added to a silica column and centrifuged at 8000 G for 30 s. The resultant flow through was discarded. The column was then washed with 600 µl of Buffer B followed by centrifugation at 8000 G for 30 s and the flow through was discarded. The column was then washed twice with 500 µl of Buffer C followed each time by centrifugation at 8000 G for 30 s and flow through was discarded. To clear any remaining buffer, the column was centrifuged for 2 min and then placed in a new 1.5 ml centrifuge tube. The RNA was captured by adding 30 µl of RNase free Mili-Q water to the column followed by centrifugation at 8000 G for 30 s. The solution containing the RNA was treated with DNase and then stored at −80°C. Additionally, to test the efficacy of the silica column against varying amounts of tissue, we performed incremental extractions from starting tissue samples that ranged from 10 mg to 60 mg (Fig. 1).

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Manufacturer protocol

Download the product protocol from Epoch Life Science Inc. for GenCatch™ Total RNA Extraction Kit below.

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