Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

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Transcription profiling of Vibrio cholerae A1552tfoX/pBAD and A1552tfoX/pBAD-tfoX

ABSTRACT: A1552tfoX/pBAD and A1552tfoX/pBAD-tfoX were diluted 1:100 from over night cultures and grown in LB liquid medium containing 0.2 % L-arabinose and 100 μg/ml ampicillin. At OD600=0.3, samples were quickly harvested in a microcentrifuge at room temperature and the bacterial pellet resuspended in Trizol reagent (GIBCO/BRL) and kept at -80:C. The aqueous phase containing the RNA was isolated according to the manufacturers manual and to which was added an equal volume of 70% ethanol. Using an RNeasy. column (Qiagen, Valencia, CA) the RNA was isolated and treated with DNase I solution. Labeling of cDNA was done as described by M. I. Voskuil et al., J. Exp. Med. 198, 705-13 (2003). Samples from A1552tfoX/pBAD were labeled with Cy3 and samples from A1552tfoX/pBAD-tfoX were labeled with Cy5. Duplicate samples from two individual experiments were hybridized to an oligonucleotide DNA microarray slide containing at least one oligonucleotide corresponding to every identified open-reading-frame of V. cholerae O1 El Tor strain N16961. Microarray slides were prehybridized (5 x SSC, 1% BSA, 0.1% SDS) for at least one hour at 42:C and hybridization was performed in 2 x SSC, 0.1% SDS, 20% formamide at 54:C over night.

ORGANISM(S): Vibrio cholerae

SUBMITTER: Janos Demeter

PROVIDER: E-SMDB-3535 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Transcription profiling of Vibrio cholerae A1552tfoX/pBAD and A1552tfoX/pBAD-tfoX

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