Guardian™ 5000 Hotplates and Stirrers

• Sample: Gram (+) positive and Gram (-) negative bacterial cells
• Yield: Up to 60 µg of RNA - (1 x 10 Escherichia coli: 40-45 µg, 1 x 10 Bacillus subtilis: 50-55 µg)
• Convenient: Includes Lysozyme and Bacteria Lysis Buffer
• Format: RNA spin columns
• Operation Time: Within 30 minutes
• Elution Volume: 50-100 µL
• Kit Storage: Dry at room temperature (15-25°C) for up to 9 months, Lysozyme should be stored at -20°C for extended periods

The rBAC Mini RNA Bacteria Kit was designed for total RNA purification from Gram (-) negative and Gram (+) positive bacteria. The provided Lysozyme and Bacteria Lysis Buffer will efficiently lyse bacterial cell walls consisting of the peptidoglycan layer. Detergents and chaotropic salt are used to further lyse cells and inactivate RNase while RNA is bound by the glass fiber matrix of the RNA spin column. Once any contaminants have been removed, using the Wash Buffer (containing ethanol), the purified total RNA is eluted by RNase-free Water and is ready for use in a variety of subsequent reactions.

Quality Control
The quality of the rBAC Mini RNA Bacteria Kit is tested on a lot-to-lot basis by isolating RNA from Escherichia coli (1x10) culture (OD600=1.3, 1 mL) harvested by centrifugation at 16,000 x g for 1 minute. 10 µL from a 50 µL eluate of purified RNA is analyzed by electrophoresis on a 0.8% agarose gel.

*Lysozyme should be stored at -20°C for extended periods. Add Lysozyme to Bacteria Lysis Buffer immediately prior to use. Once Lysozyme is mixed with Bacteria Lysis Buffer, the solution can be stored for 2 weeks at 4°C.

**Add absolute ethanol (see the bottle label for volume) to Wash Buffer then mix by shaking for a few seconds. Check the box on the bottle. Be sure and close the bottle tightly after each use to avoid ethanol evaporation.