RNeasy Mini Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	- To yield high RNA, give the column an extra wash with both RW1 and RPE buffers (3 total washes with each buffer).	- Include DNAse treatment for 15-20min. - Ensure EtOH is completely evaporated off of the column prior to elution. Adjust time from 1min to 5 min at 60`C. - Use water to elute the RNA that is warmed to ~60`C. - For purifying RNA add either 10 μl β-mercapto ethanol (β-ME) or 20 μl 2 M dithiothreitol (DTT).

Protocol tips
- To yield high RNA, give the column an extra wash with both RW1 and RPE buffers (3 total washes with each buffer).
Downstream tips
- Include DNAse treatment for 15-20min. - Ensure EtOH is completely evaporated off of the column prior to elution. Adjust time from 1min to 5 min at 60`C. - Use water to elute the RNA that is warmed to ~60`C. - For purifying RNA add either 10 μl β-mercapto ethanol (β-ME) or 20 μl 2 M dithiothreitol (DTT).

Publication protocol

Total RNA was isolated from the indicated cells using RNeasy Mini Kit (Qiagen, Valencia, CA, USA) according to the manufacturer's protocol. Reverse transcription was performed with 1 μg of total RNA using SuperScript II reverse transcriptase (Life Technologies) according to the manufacturer's instructions. The synthesized complementary DNA was amplified by PCR using specific primer sets. The PCR products were visualized by electrophoresis on agarose gels with ethidium bromide staining. GAPDH served as an internal control.

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Manufacturer protocol

Download the product protocol from Qiagen for RNeasy Mini Kit below.

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