RNeasy Mini Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	- To yield higg 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 higg 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

CAL-27 cells at a density of 1×107 cells were placed in a 75T flask and incubated without and in combination with gefitinib (10 μM) and EGCG (100 μM) for 24 h. Cells were scraped and collected by centrifugation, and total RNA was subsequently isolated using an Qiagen RNeasy Mini kit (Qiagen, Inc., Valencia, CA, USA) after being harvested as previously described (49,50). RNA quantity and purity were assessed at 260 and 280 nm using a Nanodrop ND-1000 spectrophotometer (Labtech International Ltd., East Sussex, UK).

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Manufacturer protocol

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

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