RNA isolation / purification Cells - immortalized Vero

When extracting nucleic acids from cell cultures, thorough homogenization of cells via vortexing in lysis buffer is very necessary. Choose the best RNA isolation method keeping in mind the downstream applications, generally, column-based isolations result in clean and concentrated RNA samples. Downstream applications like sequencing and cDNA synthesis require high-quality RNA, always treat the samples with DNases and check their integrity by running a gel.

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1 year ago

1 year ago by Ralf Friedmann 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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Found 3 matching solutions for this experiment

Protocol tips
- For purifying RNA add either 10 μl β-mercaptoethanol (β-ME) or 20 μl 2 M dithiothreitol (DTT).

- 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 15-20 min.

- 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.
Protocol tips
- RNA was eluted in 60 μl of AVE buffer and stored at -80°C until use.
Downstream tips
- The concentration and purity of each RNA sample were measured by Thermo Scientific NanoDrop 8000 Spectrophotometer and Agilent 2100 Bioanalyzer using the Agilent RNA 6000 Nano Kit
illustra™ RNAspin Mini Isolation Kit

GE Healthcare Life Sciences

Upstream tips
- This kit can be used for the isolation of RNA from up to 5x10^6 cells
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