E.Z.N.A.^® Bacterial RNA Isolation Kits are specially designed for isolating total cellar RNA from Gram+ and Gram- bacteria. Up to 1 x 10⁹ Bacterial cell can be processed.

Principle

The E.Z.N.A^® Bacterial RNA Kits combines the reversible nucleic acid-binding properties of HiBind^® matrix with the speed and versatility of spin column technology to yield approximately 50 -100 ug of DNA with an A²⁶⁰/A²⁸⁰ ratio of 1.8-2.0. HiBind™ matrix avidly, but reversibly, binds RNA allowing proteins and other contaminants to be removed by several wash steps. Nucleic acids are subsequently eluted with de-ionized water or low salt buffer.

Procedure

Bacterial cells are grown to log phase and harvested by centrifugation. The bacterial cell wall is weakened by lysozyme digestion. Following lysis, binding conditions are adjusted and the sample applied to an HiBind^® RNA spin-column. Two rapid wash steps remove trace salt and protein contaminants and purified RNA is eluted in water or low ionic strength buffer. Purified RNA can be directly used in downstream applications without the need for further purification.

Applications

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

• RT-PCR
• Northern Analysis
• Differential display
• Poly A⁺ RNA selection