Abstract
In order to ensure accurate gene expression analyses, it is important that the analyzed RNA truly represents in vivo transcript ratio. Generally, isolation of RNA from Gram-positive bacteria is diffi cult because these bacteria have rigid and thick cell walls. Moreover, to ensure maintenance of RNA integrity during purifi cation is diffi cult because ribonucleases (RNases) break down RNA molecules. Hence, the extraction procedures should be simple, rapid and effective. In this study we report a rapid, reproducible, high yield optimization method for the preparation of RNA from Gram-positive bacteria. The enzymatic lysis was compared with the mechanical disruption with glass beads. As cell wall disruption methods we applied lysosthaphin (37°C, 5 min, 56U) and then RNA was isolated using the TRIzol Reagent. We found that a short time of incubation is essential for the high yield recovery of bacterial RNA from Gram-positive bacteria.
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