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

The Ehrlichia chaffeensis Wakulla strain (originally provided by the Centers for Disease Control and Prevention, Atlanta, GA) was propagated in a canine macrophage cell line DH82 as described previously24 in DMEM supplemented with 10% FBS and 2% L-glutamine. At 3 d post infection, when ~95% infected of cells were infected, the DH82 cells (~5 × 106 cells) were harvested, and total RNA was extracted using an RNeasy Mini Kit (Qiagen, Valencia, CA, USA) with the optional on column DNase-digestion per 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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