The E.Z.N.A^® Yeast RNA Kits allows the convenient isolation of high-quality total RNA from a wide variety of yeast species. Up to 6 x 10⁷ log-phase culture can be processed.

Principle

The E.Z.N.A^® Yeast RNA Kits combines the reversible nucleic acid-binding properties of HiBind^® matrix with the speed and versatility of spin column technology to yield approximately 30 µg of RNA with an A260/A280 ratio of 1.7-1.9. HiBind™ matrix avidly, but reversibly, binds RNA under specific conditions allowing proteins and other contaminants to be removed. Nucleic acids are easily eluted with de-ionized water or low salt buffer.

Procedure

Yeast cells are grown to log-phase and partial spheroblasts are subsequently prepared using zymolase (or lyticase). Following rapid lysis with a highly denaturing buffer, binding conditions are adjusted and the sample applied to a HiBind^® RNA spin-column. Two efficient wash steps remove trace salt and protein contaminants and RNA is eluted in DEPC-treated water. Purified RNA can be directly used in downstream applications without the need for further purification.

Applications

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

• RT-PCR
• Hybridization Techniques

Denaturing agarose gel electrophoresis of total RNA. RNA
was isolated by E.Z.N.A.^® Yeast RNA Kit from Pichia Pastroris
culture. 15 µl of eluted RNA (total 100µl ) was loaded in each lane.