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
|- 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 during Acid-Phenol:Chloroform extraction step. This will facilitate the removal of tightly bound proteins typically associated with RNA.|
|- 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.