Vibrio cholerae Freezing Study

Question Addressed: Does gene expression in V. cholerae change when samples are frozen at -80 degrees? An in vitro grown culture of O1 Inaba ICDDR,B (strain DSM-V999 described in Nature. 2002 Jun 6;417(6889):642-5) that had been grown to exponential phase (OD600 + 0.2) was placed in a 15 ml conical and then placed in a -80 degree freezer. A portion of the starting culture was harvested prior to freezing to serve as the control/reference for downstream hybridizations. RNA was recovered from the non-frozen sample as well as by prepping material directly from frozen samples. Labeling reactions were performed in quadruplicate for each sample. A replicate experimental design type is where a series of replicates are performed to evaluate reproducibility or as a pilot study to determine the appropriate number of replicates for a subsequent experiments. replicate_design