PAXgene Tissue RNA/miRNA Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	- Add β-Mercaptoethanol (β-ME) to Buffer TM1 before use. - Buffer TM1 and Buffer TM2 may form a precipitate during storage. If necessary, warm to 37°C to dissolve - Before using carrier RNA for the first time, dissolve it (310 µg) in 1 ml RNase-free water.

Protocol tips
- Add β-Mercaptoethanol (β-ME) to Buffer TM1 before use. - Buffer TM1 and Buffer TM2 may form a precipitate during storage. If necessary, warm to 37°C to dissolve - Before using carrier RNA for the first time, dissolve it (310 µg) in 1 ml RNase-free water.

Publication protocol

Total RNA was purified from 10 mg of fresh frozen (FF), formalin or PAXgene fixed tissue or from three sections, each 10 μm thick of FFPE and PFPE tissue. For RNA isolation, the miRNeasy Mini kit (QIAGEN GmbH, Hilden, Germany) was used for FF tissue, the miRNeasy FFPE kit (QIAGEN) for formalin-fixed and FFPE tissue, and the PAXgene Tissue miRNA kit (PreAnalytiX GmbH) for PAXgene fixed and PFPE tissue. All preparations were done in triplicate with tissue samples from two different animals for each fixation method (FF, formalin-fixed, PAXgene fixed, PFPE and FFPE). RNA yield was determined by spectrophotometric absorbance (Nanodrop ND-1000 spectrophotometer, Nanodrop Technologies, Wilmington, USA) at 260 nm (A260). RNA integrity was assessed by microcapillary electrophoresis on an Agilent 2100 Bioanalyzer and analyzed with the Agilent 2100 expert software (Agilent Technologies, Waldbronn, Germany) to give RNA integrity scores (RINs).

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Manufacturer protocol

Download the product protocol from PreAnalytiX for PAXgene Tissue RNA/miRNA Kit below.

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