Found 1 discussion for this experiment
1 year ago
1 year 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-mercapto ethanol 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.|
|Total RNA from the cells was isolated using a FujiFilm
QuickGene 810 machine with QuickGene RNA cultured
cell HC kit S (FujiFilm LifeScience, FujiFilm, Tokio,
Japan). The amount and quality of isolated RNA were
measured on NanoDrop (A260/A280 and A260/A230)
(Thermo Scientific, Massachusetts, USA) and the
quality (RIN number) was checked using Agilent RNA
chips (Agilent, Colorado, USA).