General descriptionProcedures such as cDNA synthesis, expression profiling and others require separation of mRNA from the vastly more abundant rRNA and tRNA. The GenElute mRNA kits provide convenient procedures for isolating polyadenylated mRNA from previously prepared total RNA or directly from mammalian cells and tissues. For direct mRNA preparation, cells or tissues are disrupted with SDS/proteinase K digestion to release RNA and eliminate RNases. Both kit types use oligo dT30 covalently linked to 1 μm polystyrene beads to capture polyadenylated mRNA by hybridization. The polystyrene beads remain suspended during hybridization, eliminating the need for mixing or rocking, as is common for cellulose or magnetic particles. Polystyrene was also chosen because oligo(dT) polystyrene beads yield cleaner mRNA with fewer stringent washing steps than does the more commonly used oligo(dT) cellulose (2 or 3 wash steps versus 10 or more). With the GenElute kits, mRNA-bead complexes are washed on a microcentrifuge spin filter, and eluted into 10 mM Tris-HCL, pH 7.5. mRNA prepared with either kit is suitable for a variety of downstream applications such as Northern blotting, expression array or chip hybridizations and cDNA synthesis and library construction.
The GenElute mRNA Miniprep Kit provides a simple and convenient way to purify polyadenylated mRNA from previously isolated total RNA. Oligo(dT) polystyrene beads bind the poly(A)+ mRNA during a 10 minute incubation. After washing in a microspin filter to remove contaminants, the poly(A)+ mRNA is eluted in 100 mL of buffer. Purification of mRNA from total RNA, can be performed in less than 40 minutes.
The purified mRNA is ready for Northern analysis, reverse transcription and PCR, labeling for arrays, and other common applications.
Click here to view the kit protocolApplicationThe purified mRNA is ready for Northern analysis, reverse transcription and PCR, labeling for arrays, and other common applications.
Features and Benefits- Quick and convenient reagent for use in the simultaneous isolation of RNA, DNA and protein
- Performs well with large or small amounts of tissue or cells and many samples can be simultaneously extracted
- One of the most effective methods for isolating total RNA. Purifications can be completed in only one hour starting with fresh tissue or cells
- mRNA captured on oligo(dT) polystyrene beads in 10 minutes
- Oligo(dT) polystyrene beads require fewer wash steps
- Poly (A)+ mRNA isolated from previously purified total RNA in 40 minutes
Other NotesFor additional information, please see
www.sigma-aldrich.com/mrna.
Legal InformationGenElute is a trademark of Sigma-Aldrich Co. LLC
