RNeasy Plus Mini Kit

RNA isolation / purification Cells - immortalized SK-BR-3

Experiment
RNA isolation / purification Cells - immortalized SK-BR-3
Product
RNeasy Plus Mini Kit from Qiagen
Manufacturer
Qiagen

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.

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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Discussion

Discussion

4 years ago

Author: Switzerland

I am having trouble using the QiageN RNAeasy mini kit for CD34+ cells. Any tips?

I am having trouble using the QiageN RNAeasy mini kit for CD34+ cells. Any tips?

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Papers

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Manufacturer protocol

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

Download PDF Download manufacturer protocol

Videos

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