Analysis Note

Typical Sample* Cultured cells: 2 x 107Tissue: 200 mgTotal RNA: 500 µg* Larger or smaller samples may also be used.Time Required: Approximately 30 minutesProduct: Poly(A)+ RNA, in double-distilled water

Application

The mRNA Isolation Kit prepares highly purified poly (A) ⁺ RNA which may be used directly in many molecular biology applications: RT-PCR cDNA synthesis Northern blotting Northern ELISA RNase protection assay In vitro translation

Components

Lysis Buffer Streptavidin-Coated Magnetic Particles Oligo (dT)₂₀ Probe, biotin-labeled Washing Buffer Water, double-distilled, PCR Grade Storage Buffer

Features and Benefits

Save time. Isolate mRNA directly from cell lysates and tissue homogenates (isolation of total RNA not required). Accommodate a wide range of samples. Work with both small and large scale preparations of mRNA. Achieve highly effective purification. Generate mRNA that is intact and free of DNA and other RNAs. Increase safety. Eliminate the use of hazardous organic solvents.

General description

The mRNA Isolation Kit isolates highly purified mRNA from cells or tissue.Affinity purification is a versatile and highly specific technique for the purification of all classes of biomolecules utilizing differences in biological activities of chemical structures. The high selectivity of this technique results in good purification and high recovery. Often a concentrating effect is reached which enables large volumes to be conveniently processed. The mRNA Isolation Kit depends upon the affinity of the poly(A) ⁺ tail of mRNA for a biotin-labeled oligo (dT) probe. The probe can "pull" the mRNA selectively from a lysate without interacting with other RNA or DNA. Once formed, the biotinylated dT-A hybrids can be immobilized on solid surfaces that have been coated with streptavidin, and then washed free of unbound contaminants.

Other Notes

For general laboratory use.

Preparation Note

The poly (A)⁺ tail of mRNA hybridizes to a biotin-labeled oligo(dT) probe. Streptavidin-coated magnetic particles capture the biotinylated hybrids and the particles are separated using a magnetic particle separator. Contaminants are removed by washing, followed by elution of the mRNA from the particles using water.

Quality

Tested for absence of RNases according to the current quality control procedures; function tested.