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4 years ago
4 years ago by Ralf Friedmann
I am having trouble using the QiageN RNAeasy mini kit for CD34+ cells. Any tips?
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|In order to monitor the response of Dex the transfected cells, we also monitored AQP5 and GAPDH expression at the mRNA level: total RNA was extracted from a 100 μL trypsinized EpH4 cell suspension with the Quick-RNA MiniPrep Kit (Zymo Research). All steps were performed according to manufacturer's instructions. Expression changes were monitored as described above (Extraction of mRNA and analysis with qPCR), except that the SensiMix SYBR No-ROX One-step Kit (Bioline) was used for onestep qPCR.
|Three transwells from each experimental condition were harvested and pooled to isolate RNA by using the RNAqueous kit for total RNA purification from Ambion–Applied Biosystems (Austin, Tex). RNA concentration and integrity were determined by using the Agilent 2100 Bioanalyzer system and Agilent RNA 6000 Nano Chips (Agilent Technologies, Foster City, Calif).
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