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

RNA was purified from frozen tissue using the RNeasy Mini Kit (Qiagen, Copenhagen, Denmark) according to manufacturer's instructions. For each batch of purification, a maximum of 30 mg of tissue was homogenized at 25 Hz for 3 min using a Tissuelyser II machine (Qiagen) and 5 mm Stainless Steel Beads (Qiagen). Homogenized tissue was purified using RNeasy Mini columns, and subsequent to purification, on‐column DNase digestion of the RNA for elimination of DNA contamination was performed (RNase‐Free DNase Set, Qiagen). RNA was purified from FFPE samples/formalin‐fixed tissues using the RNeasy FFPE Kit (Qiagen). Five sections of 10 μm were pooled and deparaffinated in deparaffination Solution (Qiagen) and RNA was subsequently purified according to manufacturer's instructions for optimized removal of genomic DNA contamination. Quantification of total RNA was measured using a Thermo Scientific NanoDrop 1000™ (Thermo Scientific, Hvidovre, Denmark) and Thermo Scientific Software ND.1000 V3.5.2.

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Manufacturer protocol

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

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