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Oligotex Suspension

For mRNA purification
  • High recovery of pure mRNA in as little as 30 minutes
  • No oligo-dT cellulose or ethanol precipitation
  • Flexibility for use with widely varying amounts of starting RNA
Each Oligotex Kit includes Oligotex resin for binding poly A+ mRNA, and spin columns for washing and eluting bound mRNA. Oligotex mRNA Kits enable purification of mRNA from total RNA and cleanup of poly A+ in vitro transcripts. Oligotex Direct mRNA Kits enable purification of mRNA from cells and tissues. Oligotex Suspension provides additional Oligotex resin for use with Oligotex Kits.
Cat No./ID: 79000
Oligotex Suspension (0.5 ml)
$1,159.00
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0.5 ml for mRNA isolation from up to 8 mg of total RNA
The Oligotex Suspension is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.
Performance

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").

Principle

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.

Procedure

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.

Applications

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

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Kit Handbooks (1)
For Purification of poly A+ RNA from total RNA, purification of poly A+ RNA directly, from cultured cells or tissues, and purification of polyadenylated in vitro transcripts 
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User-Developed Protocols (2)
The procedure has been used successfully by customers for regeneration of Oligotex resin from spin columns following elution of poly A+ mRNA.
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The procedure has been successfully used for isolation of mRNA from adipose tissue by customers. Because of the nature of adipose tissue it cannot be used immediately in Oligotex Direct mRNA Protocols; simple centrifugation, however, eliminates the problem, and the procedure works fine.
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