TriPure Isolation Reagent

Protocol tips

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

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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Manufacturer protocol

Download the product protocol from Sigma-Aldrich for TriPure Isolation Reagent below.

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