miRNeasy Mini kit

RNA isolation / purification Tissue - Rat Nose

Experiment

RNA isolation / purification Tissue - Rat Nose

Product

miRNeasy Mini kit from Qiagen

Manufacturer

Qiagen

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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

Nasal epithelial tissue samples were disrupted and homogenized using a TissueRuptor (Qiagen) and RNA isolated using the miRNeasy kit (Qiagen). For the WBC, samples were homogenized in TRIzol (Life Technologies) and RNA isolated according to the standard TRIzol protocol. The BM samples were filtered through 70-μm nylon mesh filters (Fischer Scientific, Waltham, Massachusetts) to remove bone fragments. Filtered BM samples were then homogenized in TRIzol (Life Technologies) and RNA isolated according to the standard TRIzol protocol. Extracted RNA was quantified with a Nanodrop 1000 spectrophotometer (Thermo Scientific, Waltham, Massachusetts) and its integrity verified with a 2100 Bioanalyzer (Agilent Technologies, Santa Clara, California) using both the Nano and Small RNA kits. The quality control incorporated RNA integrity number assessment and electrophoretogram peak assessment of small RNA molecules.

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Reviews

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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?

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Discussion

5 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 Mini kit below.

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