RNeasy Mini Kit

RNA isolation / purification Cells - immortalized MRC5VA

Experiment
RNA isolation / purification Cells - immortalized MRC5VA
Product
RNeasy Mini Kit from Qiagen
Manufacturer
Qiagen

Protocol tips

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.

Publication protocol

total RNA was extracted using RNeasy kits (Qiagen) with on-column DNase digestion. Fractionated protein extracts were prepared by sequential extraction in NP-40 buffer (0.5% NP-40, 25 mM Tris pH7.9, 150 mM NaCl), Dignam buffer C (20 mM HEPES pH7.9, 25% glycerol, 0.42 mM NaCl, 1.5 mM MgCl, 0.2 mM EDTA) and Laemmli buffer (50 mM Tris pH6.8, 2% SDS, 10% glycerol). Alternatively, whole cell protein extracts were prepared by direct addition to cells of hot Laemmli buffer and incubation at 110°C for 10mins with intermittent vortexing. The protein concentration of each sample was determined after suitable dilution using a Bicinchoninic Acid (BCA) assay (Thermo Scientific).

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Discussion

Discussion

1 year 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 Mini Kit below.

Download PDF Download manufacturer protocol

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