Use of a Dental Amalgamator to Extract RNA from the Gram-Positive Bacterium Streptococcus agalactiae

摘要

The isolation of RNA from Streptococcus and other gram-positive bacteria has proven difficult because these organisms possess thick, rigid cell walls that prevent lysis. Generally, two methods are used to disrupt the cell wall: (i) degradative enzymesthat digest and weaken the cell wall or (ii) mechanical agitation by mixing bacterial cells with zirconia/silica beads in a high-speed reciprocating shaker. Although enzymatic digestion is effective, it usually takes several minutes to complete. Duringthis time, RNA processing/degradation can occur, resulting in poor quality and yield. Reciprocating shakers are a fast and effective means of lysing bacteria, but these instruments are often expensive. We describe the isolation of RNA from S. agalactiaeusing an inexpensive dental amalgamator as an oscillating shaker to lyse these bacteria.