TriPure Isolation Reagent

RNA isolation / purification Tissue - Mouse Liver

Experiment
RNA isolation / purification Tissue - Mouse Liver
Product
TriPure Isolation Reagent from Sigma-Aldrich
Manufacturer
Sigma-Aldrich

Protocol tips

Upstream tips
- Process tissues immediately and freeze at -60c or below for Low RNA degradation
Protocol tips
- Adding too little reagent can lead to contamination with DNA

Publication protocol

Firstly, liquid nitrogen was added and the samples were ground separately. Then, either 1 ml Trizol reagent (Invitrogen, USA) or 1 ml TriPure (Roche, USA) was added. Upon completion of the harvest procedure, the homogenates were transferred to empty RNase free falcon tubes stored on ice. To prevent microbial contamination and subsequent RNase contamination, disposable gloves were always used and good sterile technique and methods were practiced when handling samples. Also, the gloves were changed frequently. Then, ethanol (250 μl), isopropanol (500μl), chloroform (200 μl) were added respectively. Subsequently, once b–mercaptoethanol was added to the homogenate to remove the RNase. Ultimately, the quality of RNA was checked on agarose gel electro-phoresis to observe the 5S, 18S and 28S bands.

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Discussion

Discussion

2 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

3 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 Sigma-Aldrich for TriPure Isolation Reagent below.

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