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

otal RNA was isolated using TRIzol (Invitrogen, Mulgrave, VIC, Australia) and the RNeasy Mini Kit (Qiagen, Valencia, CA, USA) from 12 NMC cell lines (11060, 14169, 8645, RPMI2650, TC797, TY82, 10326, HCC2429, PER-403, PER-624, PER-704, P896-CL) and the P896-FB fibroblast line according to the manufacturers’ protocols, with QC performed using the Bioanalyzer 2100 (Agilent Technologies Australia, Mulgrave VIC).

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Manufacturer protocol

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

Download manufacturer protocol

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