Oligotex mRNA Midi Kit

Oligotex mRNA kits are available for isolation of mRNA from less than 0.25 mg up to 1 mg of total RNA starting material (see Table).

Poly A⁺ mRNA purified with Oligotex resin is suitable for RT-PCR (see figure "RT-PCR of beta-actin mRNA") and poly A⁺ mRNA isolated from total RNA with Oligotex resin has been used for Northern, dot, or slot blotting (see figure "Isolation of mRNA from total RNA").

Oligotex mRNA Kits contain spin columns and all necessary reagents and buffers for isolation of pure poly A⁺ mRNA from total RNA preparations or cleanup of in vitro transcripts. Oligotex resin consists of polystyrene–latex particles of uniform size (1.1 µm diameter) and a perfect spherical shape (see figure "Oligotex particles and oligo-dT cellulose") with dC₁₀T₃₀ oligonucleotides covalently linked to the surface. The size, composition, and surface structure of Oligotex particles have been optimized for uniform dispersion and minimal centrifugation time. The particles form a stable suspension that provides a large surface area for rapid and efficient binding of polyadenylic acids.

The high capacity and accuracy of hybridization provides superior purification of poly A⁺ mRNA through fast and efficient hybridization. mRNA recoveries are consistently greater than 90%. The Oligotex purification procedure minimizes loss of poly A⁺ RNA, eliminates the risk of degradation by RNases, and requires minimal hands-on time. This makes it ideal for simultaneous handling of multiple samples. Poly A⁺ mRNA is immediately ready for all standard applications, as well as for sensitive applications such as expression array, differential display, cDNA library construction, microinjection, and SAGE technology. Oligotex Suspension is also available separately.

Optimized Oligotex purification procedures are convenient and straightforward (see flowchart "Oligotex mRNA procedure"). Total RNA preparations are incubated with Oligotex resin, and Oligotex:mRNA complexes are collected by a brief centrifugation step. Spin columns, supplied in the Oligotex Kits, provide the most convenient handling, and optional batch protocols are also provided for certain applications. After washing, the mRNA is eluted in a small volume of Tris buffer or water. Alcohol precipitation is not required.

Poly A⁺ mRNA purified with Oligotex resin is ready for use in any downstream application, including:

• RT-PCR
• cDNA synthesis
• cDNA library construction
• Subtractive hybridization
• In vitro translation