A protocol for the isolation of high-purity RNA from gram-positive and gram-negative bacteria was developed for use with the Agilent Total RNA Isolation Mini Kit.
The document presents an optimized protocol for isolating total RNA from gram-positive and gram-negative bacteria using the Agilent Total RNA Isolation Mini Kit. The method utilizes a phenol-free spin-column approach with a unique prefiltration column to remove cellular contaminants, including genomic DNA, often eliminating the need for subsequent DNase treatment.
Gram-positive cells should be digested for 20-40 minutes.
The recommended concentration is 3 mg/mL.
Centrifuge for 3 minutes at 16,000 x g.