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The total RNA purification method for difficult plant tissues
Detailed Product Description
Many kits, reagents and lab protocols work for isolating total RNA from plant tissues like young tomato leaves, Arabidopsis leaves and soybean leaves. However, when older tissue, tissue from stressed plants or tissues high in secondary metabolites are used, these methods falter or fail in isolating quality total RNA.
Other methods used to isolate total RNA from plant tissues:
- Acid-guanidine thiocyanate>phenol-chloroform extraction (TRIzol® and TRI® Reagent)
- salting out and precipitation
- kits using silica spin columns (ex. RNeasy®)
- magnetic beads
The Spectrum Plant Total RNA Kit performs similarly to other methods with young leaf tissue from tomato and Arabidopsis. However, the kit outperforms nearly all other conventional protocols with more challenging tissue samples, such as pine needles or cotton leaves.
The diversity of secondary metabolites in plants is a rich and invaluable resource, yet data and experience show reduced efficiency of nucleic acid purification protocols in the presence of high levels of polyphenolics, polysaccharides, tannins, isoflavanoids, waxes, oils and other metabolic compounds. The Spectrum Plant Total RNA Kit was specifically developed to optimally perform in the presence of secondary metabolites. This optimal performance was achieved through research that identified the mode of failure in common kit-based total RNA purification protocols and development that eliminated the mode of failure.
In various column-based kit protocols, experimental data suggested that metabolic compounds were affecting the interaction between the RNA and the binding substrate and thus reducing the binding column’s functionality. These compounds were also carrying through the entire purification process, resulting in contamination of the eluted RNA. To correct this widely observed shortcoming, a chemistry novel in RNA purification protocols was reengineered by Sigma and made suitable for this application. This chemistry is highly effective in removing nearly all secondary metabolites before the RNA is bound to the binding substrate or even washed.
The result of this redevelopment is a total RNA isolation kit for plants that is quick, convenient and capable of purifying high quality of total RNA in the presence of secondary metabolites.
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