High Pure RNA Isolation Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
- To decrease RNase activity,process starting material immeditely or store at 80C. - To increase nucleic yield or purity store all buffer at +15C to +25C.	- To decrease RNase activity,use eluted RNA directly in downstream procedure or store at -80C immediately

Upstream tips
- To decrease RNase activity,process starting material immeditely or store at 80C. - To increase nucleic yield or purity store all buffer at +15C to +25C.
Protocol tips
- To decrease RNase activity,use eluted RNA directly in downstream procedure or store at -80C immediately

Publication protocol

Kidney tissues snap frozen and stored at -80°C were used for RNA isolation. Tissue piece (100 mg) was transferred to a DNA/RNA-free round bottom tube containing CHAPS buffer (Cat. No. 90436, Millipore Co., CA) and gently homogenized (TEKMAR Tissuemizer Co., Cincinnati, OH) for 10–20 s (seconds). The homogenized tissue was kept in ice for 20 min followed by centrifugation (12,000 x g for 20 min). The supernatant (200 μL) was mixed with 400 μL of RNA lysis buffer (High Pure RNA Isolation kit, Cat. No.11 828 665001, Roche Diagnostics, Mannheim, Germany) and the procedure was continued as per manufacturer’s instructions. The isolated RNA was assessed for purity and quantity by spectrophotometry (Nanodrop ND-100, Nanodrop technologies, Inc., Wilmington, DE). A ratio >2 OD @ 260 nm/280 nm was considered high quality RNA for further studies.

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Manufacturer protocol

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