The GenElute™ Direct mRNA Miniprep kit provides a convenient format to isolate polyadenylated mRNA directly from mammalian cells and tissues. The direct mRNA isolation procedure is based on that of Badley. Up to 107 mammalian cells or 40 mg tissue are lysed and homogenized, either with the filtration columns provided or with
a mechanical homogenizer. RNase is eliminated during a 10 minute proteinase K digestion. Sodium chloride is added, and polyadenylated RNA is captured on oligo(dT) polystyrene beads during a 10 minute incubation. For further enrichment, RNA may be released from the beads into fresh lysis solution and recaptured with the original beads. After 3 washes in a spin column, purified mRNA is eluted in 100 μl of 10 mM Tris-HCl, pH 7.4.
GenElute™ Direct mRNA Miniprep Kit has been used to isolate mRNA from various tissues and cells.
The GenElute Direct Kit mRNA kit provides convenient procedures for isolating polyadenylated mRNA from previously prepared total RNA or directly from mammalian cells and tissues.
Features and Benefits
- mRNA captured on oligo(dT) polystyrene beads in 10 minutes, with no mixing or rocking (Fig. 1)
- Poly (A)+ mRNA isolated from total RNA in 40 minutes (Fig. 2) or 60 minutes directly from cells and tissues (Fig. 3)
- Oligo(dT) polystyrene beads require fewer wash steps
Use for isolating mRNA directly from mammalian cells or tissues.
For additional information, please see www.sigma-aldrich.com/mrna.
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