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).
|- To reduce RNA degradation do not wash cells before the addition of RNAzol® RT.
- To reduce DNA contamination, use sufficient amount of RNAzol® RT based on the area of the culture dish and not on cell number.
|To reduce DNA contamination extend the incubation time to 15 minutes before sedimentation.|