GenElute™ Mammalian Total RNA Miniprep Kit

Description

• General description: Sigma′s GenElute Mammalian Total RNA Miniprep Kit provides a simple and convenient way to isolate total RNA from mammalian cells and tissues. Protocols are provided for cells, tissues, and fibrous tissues. These protocols differ in their cell lysis and disruption conditions. Once the RNA is bound to the GenElute Binding Column, the purification procedure is the same for all starting materials. For fibrous tissues the kit must be used with proteinase K (Cat. No. P4850) to ensure effective cell disruption. The kit combines the advantages of a silica-based system with a microspin format and eliminates the need for cesium chloride gradients, alcohol precipitation, and hazardous organic compounds such as phenol and chloroform. Cells or tissues are lysed and homogenized in a buffer containing guanidine thiocyanate to ensure thoroughdenaturation of macromolecules and inactivation of RNases. Addition of ethanol causes RNA to bind when the lysate is spun through a silica membrane in a microcentrifuge tube. After washing to remove contaminants, RNA is eluted in 50-100 μL of Elution Solution. Up to 150 μg of total RNA can be isolated in less than 30 minutes. If all traces of DNA contamination must be eliminated, further treatment with DNase I is recommended. DNase I digestion can be performed while the RNA is bound to the GenElute Binding Column using the On-Column DNase I Digestion Set (DNASE10 and DNASE70). Alternatively, for more stringent removal of contaminating DNA, the final RNA preparation can be treated with Amplification Grade DNase I (AMPD1).
• Application: GenElute™ Mammalian Total RNA Miniprep Kit has been used to extract total RNA from tissues and cells.[1][2][3]
• Features and Benefits: Purifies total RNA from up to 107 cells or 40 mg of tissue per prepYields up to 150 μg of pure, concentrated total RNA per prepRecover RNA from as few as 100 cellsSimple and efficient–12 to 18 preps in 30 minutesFaster than gravity flow anion exchange methodsNo cumbersome steps associated with resins and magnetic slurries40% more purifications per kit than the leading supplier
• Other Notes: The GenElute™ Mammalian Total RNA Purification Kit combines silica-membrane technology with a convenient spin column format for a rapid bind, wash, and elute method to prepare high quality total RNA.
• Principle: Samples are lysed and homogenized in guanidine thiocyanate and 2-mercaptoethanol to release RNA and inactivate RNases. Lysates are spun through a filtration column to remove cellular debris and shear DNA. The filtrate is then applied to a high capacity silica column to bind total RNA, followed by washing and elution. Up to 150 μg of total RNA can be recovered per prep in 100 μl of water. The purified RNA is ready for Northern blots (Fig. 1), RT-PCR (Fig. 2) and other common applications.
• Legal Information: GenElute is a trademark of Sigma-Aldrich Co. LLC