Found 1 discussion for this experiment
2 years ago
2 years ago by Ralf Friedmann
I am having trouble using the QiageN RNAeasy mini kit for CD34+ cells. Any tips?
Found 3 matching solutions for this experiment
|- To yield high RNA, give the column an extra wash with both RW1 and RPE buffers (3 total washes with each buffer).|
|- 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.
- For purifying RNA add either 10 μl β-mercapto ethanol (β-ME) or 20 μl 2 M dithiothreitol (DTT).
|- When processing <500 cells or <2 µg tissue, carrier RNA may be added to the lysate before homogenization.
- Foaming can be reduced by adding Reagent DX at a final concentration of 0.5% (v/v) before disruption and homogenization.
- Add 4 volumes of ethanol (96–100%) to Buffer RPE for a working solution.