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

Cells were collected from 226L-U19, HB4a, MCF10a, MCF7, T47D and SKBR3 cell lines cultured in monolayer (pre-incubated with MS media for 12 hrs) and from viable AR cells. RNA was extracted using the RNeasy® Plus Mini (Qiagen) following the manufacturers protocol. In addition to the gDNA eliminator spin column provided in the RNeasy kit, an on-column DNase digestion was performed (RNase-Free DNase Set; Qiagen) to ensure maximum removal of DNA. RNA was eluted and the concentration measured using a GeneQuant machine (Amersham Biosciences). RNA integrity was assessed by microanalysis (Agilent Bioanalyser) and amplified using the WT-Ovation™ Pico RNA Amplification System (NuGEN) following the manufacturers protocol which employs SPI™ amplification (linear isothermal DNA amplification process) to amplify the RNA about 15,000-fold.

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Manufacturer protocol

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

Download manufacturer protocol

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