NucleoSpin® RNA

Protocol tips

Upstream tips	Protocol tips	Downstream tips
- Aliquot rDNase and store at -20 °C.	- Dry the columns very well after the ethanol wash by adding an additional 2' >10000 rpm centrifuge with no buffer.

Upstream tips
- Aliquot rDNase and store at -20 °C.
Protocol tips
- Dry the columns very well after the ethanol wash by adding an additional 2' >10000 rpm centrifuge with no buffer.

Publication protocol

Quadruplicates samples of 6×104 cells (BxPC-3, MIA PACA-2 and CHO-WT) were seeded in 12-well plates and initially cultured for 48 h. Cell total-RNA was extracted using Nucleospin RNA II (Macherey-Nagel, Duren, Germany) with on-column DNA digestion, according to manufacturer's instructions. TRIzol LS (Life Technologies Corporation, Carlsbad, CA, USA) was used for the homogenization of the human tissue sample in a Mini-BeadBeater-1 (Biospec Products, Coventry, UK). The RNA-containing aqueous phase was isolated after Chloroform phase separation followed by an Isopropanol nucleic acid-precipitation step. The resulting RNA pellet was further processed with the Nucleospin RNA II kit as above. RNA eluates' concentration and purity were determined with NanoDrop 2000 (Thermoscientific, Open Biosystems, Lafayette, CO, USA).

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Manufacturer protocol

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