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 extracted from the cells using the RNeasy Mini kit (Qiagen Inc., Valencia, CA, USA) as described previously11. Briefly, the cells were lysed and homogenized in RLT buffer supplemented with 10 µl/ml mercaptoethanol (Sigma-Aldrich, Munich, Germany). The lysate was homogenized using a syringe and a 20-G needle. The sample was placed in a silica column followed by washing and eluting with RNase-free water. The RNA concentration was measured using an ultraviolet spectrophotometry at 260 nm, and the purity and integrity were evaluated using the A260/A280 ratio.

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Manufacturer protocol

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

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