miRNeasy FFPE Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
- To allow complete penetration by formalin, use tissue samples less than 5 mm thick	- Perform all centrifugation steps using a microcentrifuge placed at 15–25°C.	- Do not exceed fixation time of 24 hours as it results in poor performance in downstream assay.

Upstream tips
- To allow complete penetration by formalin, use tissue samples less than 5 mm thick
Protocol tips
- Perform all centrifugation steps using a microcentrifuge placed at 15–25°C.
Downstream tips
- Do not exceed fixation time of 24 hours as it results in poor performance in downstream assay.

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 Qiagen for miRNeasy FFPE Kit below.

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