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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 3 matching solutions for this experiment
|- For RNA to dissolve better and to yield 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.|
|mRNA and protein was extracted from monolayers of HEKn and HaCaT cells using AllPrep Protein/RNA Isolation Kit according to manufacturer’s protocol (QIAGEN GmBH, Hilden Germany), quantified using the NanoDrop Lite Spectrophotometer (ThermoScientific, Wilmington, USA) and expression of differentiation markers Keratin 14 (K14), Keratin 10 (K10) and Involucrin (Inv) assessed by qRT-PCR and Western blot.|