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.

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.

Publication protocol

Total RNA was isolated from cells using the RNeasy Mini Kit (Qiagen) according to the manufacturer’s protocol. RNA concentration was determined by absorbance at A260 and RNA purity was determined by A260/A280. cDNA templates were generated using 1 μg of total RNA in 20 μl reactions using High-Capacity cDNA Reverse Transcription Kits from Applied Biosystems, Inc (Foster City, California) according to the manufacturer’s protocol. qPCR was performed in a 20 μl reactions with 4 ng of cDNA template and 900 nM primer using a SsoFast EvaGreen supermix (Bio-Rad Laboratories Inc, Hercules, California) according to the manufacturer’s protocol.

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Manufacturer protocol

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

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