RNeasy Plus Mini Kit

Protocol tips

Upstream tips	Protocol tips	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.

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

Total RNA was prepared using the RNeasy Plus Mini Kit (Qiagen) from independent cultures of cells that have been seeded at a density of 750,000 cells in a 10-cm dish and grown for 2 d in F12 media with 10% FBS without selection to 50–75% confluency at the time of harvest. RNA quality was confirmed using capillary electrophoresis (Bioanalyzer 2100; Agilent), amplified, and hybridized to an Agilent G3 Human GE 8×60K dual-color microarray by the University of North Carolina Lineberger Comprehensive Cancer Center Genomics and Bioinformatics Core Facility. Microarrays were scanned using an Axon 4200 Scanner (Molecular Devices), normalized using GenePix 5.0 software (Molecular Devices), and analyzed using the Partek Genomics Suite 6.5 software. ANOVA and t test analysis were performed using Genomics Suite 6.5 software with an FDR cutoff of P < 0.05. Clustering of the significantly changed gene set was also performed with Genomics Suite 6.5 software.

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Manufacturer protocol

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

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