PAXgene Tissue RNA/miRNA Kit

RNA isolation / purification Tissue - Rat Heart

Experiment
RNA isolation / purification Tissue - Rat Heart
Product
PAXgene Tissue RNA/miRNA Kit from PreAnalytiX
Manufacturer
PreAnalytiX

Protocol tips

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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Discussion

Discussion

4 years 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

4 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 PreAnalytiX for PAXgene Tissue RNA/miRNA Kit below.

Download PDF Download manufacturer protocol

Videos

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