E.Z.N.A.^® Total RNA Midiprep Kit provides a rapid and easy method for the isolation of up to 1mg of total RNA from cultured eukaryotic cells, tissues, or bacteria. The kit allows the simultaneous processing of single or multiple samples in less than 30 minutes Normally, up to 1 x 108 eukaryotic cells or 200 mg tissue can be used in a single experiment.

Principle

By using the HiBind® Midi spin-column technology, this E.Z.N.A^® Total RNA Midiprep Kit greatly reduces the processing time when compared to traditional phenol methods. The E.Z.N.A.^® Total RNA Midiprep Kit uses the reversible binding properties of the HiBind^® matrix, a new silica-based material. A specially formulated high salt buffer system allows more than 100 µg of RNA molecules greater than 200 bases to bind to the matrix.

Procedure

Cells or tissues are combined with TRK Lysis Buffer and homogenized to efficiently extract RNA. Equal volumes of ethanol are added to the to the lysate to precipitate RNA and facilitate binding to the spin column. Samples are applied to HiBind^® RNA Midi spin-column and subjected to a wash step with RNA Wash Buffer I. At this point an optional on column DNase I digestion reaction can be applied to if high purity RNA is required. These processes are followed by additional washes with RNA Wash Buffer II. After completion of the wash steps RNA is eluted with DEPC-treated water.

Applications

RNA purified using the E.Z.N.A.^® Total RNA Midiprep Kit can be used in applications such as :

• RT-PCR
• Northern Blotting
• Nuclease Protection
• In vitro translation