RNeasy Micro Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
- This kit can be used for the isolation of RNA from up to 5x10^5 cells, or up to 5 mg of tissue.		- Always blank the spectrophotometer with your elution buffer. - When measuring 260 nm absorbance, a unit of 1 on this scale equals to 40 ng of RNA.

Upstream tips
- This kit can be used for the isolation of RNA from up to 5x10^5 cells, or up to 5 mg of tissue.
Downstream tips
- Always blank the spectrophotometer with your elution buffer. - When measuring 260 nm absorbance, a unit of 1 on this scale equals to 40 ng of RNA.

Publication protocol

Total RNA was isolated using the RNeasy Micro Kit (Qiagen, Hilden, Germany) according to the manufacturer's instructions, with minor modifications. For 0 and RA treated cells, 1.4 ml was harvested from each flask, centrifuged at 420xg for 5 min in RNAse-free tubes, supernatants removed and 1.4 ml of RLT-buffer containing ß-Mercaptoethanol was added. For TPA treated cells, dead cells were decanted from the Petri dish and 1.75 ml RLT-buffer containing ß-Mercaptoethanol was added and cells scraped (yielding approximately the same cell concentration as the 0 and RA treated cells). Each of the 4 replicate samples was individually lysed by vigorous vortexing, immediately frozen in dry ice and stored at −80 C°. Subsequently, each thawed lysate was homogenized using QIAshredder spin columns (Qiagen, Hilden, Germany). One volume of 70% EtOH was added, the RNA bound to a RNeasy MinElute spin column and genomic DNA digested by DNAse I. Afterwards, DNase I and residual buffer traces were removed by subsequent washing steps and the RNA eluted in 14 µl nuclease-free water (Ambion, ThermoFisher Scientific, Waltham, MA). Integrity and purity of RNA was ensured using a Bioanalyzer 2100 (Agilent, Santa Clara).

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Manufacturer protocol

Download the product protocol from Qiagen for RNeasy Micro Kit below.

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