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

RNA was extracted from cultured cells using an RNeasy Mini Kit (Qiagen Japan), and 1 μg of total RNA was converted to cDNA with the High-Capacity cDNA Reverse Transcription Kit (Applied Biosystems). The human TRAIL receptors (R1 and R2), TRAIL decoy receptors (DcR1 and DcR2), and β-actin were amplified by SYBR Premix Ex Taq (Takara Bio). All procedures were carried out according to the manufacturer's instructions.

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Manufacturer protocol

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

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