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Transcriptional profiling of wild-type, mglA point mutant, and fevR (FTT0383) knockout bacteria

ABSTRACT: Time courses and RNA isolation. Overnight cultures of F. novicida strain U112, GB2, and fevR ko (383) were grown in TSB 0.2% cysteine at 37C with shaking and subcultured to an OD600 of 0.01 in 200ml. Samples were taken throughout the growth curve for RNA isolation and to determine the OD600. Five samples were taken during the growth curve at 1, 3, 5, and 7.5h and 10h for characterization of single mutants or at 2, 4, 6, and 8h in gro::fevR bypass experiments. RNA was harvested and a common reference was created from each individual sample. 1ug of either the Sample, or Reference was reverse transcribed and labeled with either Cy5 (samples) or Cy3 (reference) and hybridized to a Francisella microarray previously described in Brotcke, Weiss, et al, Infection and Immunity, 2006. Groups of assays that are related as part of a time series. Age: Red channel represents 1ug of RNA from an individual timepoint Starting OD: Wt, GB2, and FTT0383 (fevR) ko were subbed to an OD of 0.01 in TSB + 0.2% cysteine in 200ml. Keywords: time_series_design

ORGANISM(S): Francisella tularensis

PROVIDER: GSE11554 | GEO | 2009/01/06

SECONDARY ACCESSION(S): PRJNA106283

REPOSITORIES: GEO

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Transcriptional profiling of wild-type, mglA point mutant, and fevR (FTT0383) knockout bacteria

Project description:Time courses and RNA isolation. Overnight cultures of F. novicida strain U112, GB2, and fevR ko (383) were grown in TSB 0.2% cysteine at 37C with shaking and subcultured to an OD600 of 0.01 in 200ml. Samples were taken throughout the growth curve for RNA isolation and to determine the OD600. Five samples were taken during the growth curve at 1, 3, 5, and 7.5h and 10h for characterization of single mutants or at 2, 4, 6, and 8h in gro::fevR bypass experiments. RNA was harvested and a common reference was created from each individual sample. 1ug of either the Sample, or Reference was reverse transcribed and labeled with either Cy5 (samples) or Cy3 (reference) and hybridized to a Francisella microarray previously described in Brotcke, Weiss, et al, Infection and Immunity, 2006. Groups of assays that are related as part of a time series. Age: Red channel represents 1ug of RNA from an individual timepoint Starting OD: Wt, GB2, and FTT0383 (fevR) ko were subbed to an OD of 0.01 in TSB + 0.2% cysteine in 200ml. Keywords: time_series_design Computed

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