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 4 matching solutions for this experiment
|- For RNA to dissolve better and to yeild high levels, preheat elution buffer at 95C.
- Addition of β-mercaptoethanol in lysis buffer can be skipped because the phenol will do the job of nuclease inactivation.
- To capture more amount of mRNA, modify the amount of lysis buffer :ethanol (from 1:1 to 1:1.5) during the binding step.
- Do vortex for 2-plus minutes. This will facilitate the removal of tightly bound proteins typically associated with RNA.
|- 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.|