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

For transcriptome analysis, RNA from DAOY cells was extracted by using Trizol (Life Technologies). Briefly, cells were washed with phosphate buffer saline (PBS) and lysed in 1 ml Trizol. Chloroform (0.2 ml) was added, and the samples were mixed intensively and incubated for 5 min at room temperature. The upper aqueous phase was transferred to a new tube after centrifugation (12 000 g, 15 min, 4 °C) and mixed with 0.5 ml of isopropanol. After incubation at 4 °C for 10 min, the precipitated RNA was pelleted by centrifugation (12 000 g/15 min/4 °C) and washed with 75 % ethanol. The RNA pellet was dissolved in 20 µl of RNase-free water. The RNA was stored at −80 °C until further use.

For qRT-PCR analysis, total cellular RNA was isolated using the NucleoSpin RNA Plus kit (Macherey-Nagel).

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Manufacturer protocol

Download the product protocol from Macherey Nagel for NucleoSpin® RNA below.

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