GenElute(TM) Direct mRNA Miniprep Kits (C15-1356-039)

General description

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.

Application

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

Other Notes

Use for isolating mRNA directly from mammalian cells or tissues.

For additional information, please see www.sigma-aldrich.com/mrna.

Legal Information

GenElute is a trademark of Sigma-Aldrich Co. LLC