miRNeasy FFPE Kit

RNA isolation / purification Tissue - Rat Spleen

Experiment
RNA isolation / purification Tissue - Rat Spleen
Product
miRNeasy FFPE Kit from Qiagen
Manufacturer
Qiagen

Protocol tips

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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Discussion

Discussion

1 year ago

Author: Paul G. Macon United States

RNA isolation from tissue

How do I extract RNA from animal tissue without using liquid nitrogen? I tried the RNA extraction by using the TRIzol reagent and I homogenize the tissue using polytron homogenizer at room temperature for 30secs is this correct?

Discussion

2 years ago

Author: Aaron Stege Netherlands

Problem in phase separation after using serum/plasma kit

I used a serum/plasma kit for my serum samples. After the phase separation the samples should have 3 phases: a colourless aqueous phase, a white interphase and a red organic phase. However, in some of my samples there was no aqueous phase unless I wait for an extended period of time. How can I circumvent this problem?

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Papers

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Manufacturer protocol

Download the product protocol from Qiagen for miRNeasy FFPE Kit below.

Download PDF Download manufacturer protocol

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