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. - For purifying RNA add either 10 μl β-mercapto ethanol (β-ME) or 20 μl 2 M dithiothreitol (DTT).

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. - For purifying RNA add either 10 μl β-mercapto ethanol (β-ME) or 20 μl 2 M dithiothreitol (DTT).

Publication protocol

Total RNA was extracted from cells using an RNeasy Mini kit (Qiagen, Tokyo, Japan) and reverse transcribed using a cDNA reverse transcription kit (Applied Biosystems, Foster City, CA, USA). qRT-PCR analysis was performed on an ABI PRISM 7700 Sequence Detection System using SYBR-Green PCR Master Mix (both from Applied Biosystems) and the following cycling conditions: 95°C for 10 min, followed by 50 cycles of 95°C for 15 sec and 60°C for 1 min

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Manufacturer protocol

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

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