Illumina Genome Analyzer IIx sequencing; Genomic mapping of cAMP receptor protein binding in relation to transcriptional start sites in Mycobacterium tuberculosis
Genomic mapping of cAMP receptor protein binding in relation to transcriptional start sites in Mycobacterium tuberculosis
M. tuberculosis strains H37Rv (wild type) and M. tuberculosis Deltacrp were grown in Dubos broth containing 0.05% (v/v) Tween and supplemented with 0.5% (v/v) glycerol and 4% albumin. To monitor the response to nutrient starvation, M. tuberculosis was grown to mid-exponential phase (OD600 0.6), the cells pelleted and washed twice in PBS supplemented with 0.025% tyloxapol. Cells were then resuspended in 100 milliliters PBS plus tyloxapol and incubated without shaking at 37 degrees. For ChIPseq, M. tuberculosis cells were grown in roller bottles to mid-exponential phase and formaldehyde was added to a final concentration of 1 percent. After 10 minutes of incubation, glycine was added  and cross-linked cells were harvested. RNA was extracted from exponentially growing wild type and Deltacrp M. tuberculosis H37Rv as previously described (E-MTAB-1616). Samples were sequenced on an illumina Genome Analyzer IIx as single end reads
Experimental Factor: growth condition
exponential
Experimental Factor: genotype
wild type