Vibrio cholerae wt; exponential culture sample; LB
This sample was intended for Chromosome Conformation Capture (3C): 3C libraries were built, sequenced and analyzed as previously described by Marbouty et al. (2015) PMID: 26295962; doi: 10.1016/j.molcel.2015.07.020. Construction of 3C libraries: Briefly, cells were grown in 100 ml of LB medium at 30°C until a final concentration of ~1-2 x 107 cells/ml. Cells were then crosslinked using fresh formaldehyde and stored at -80°C. Thawed cells were lyzed and the cross-linked DNA digested with HpaII. The insoluble fraction of the sample was pelleted. Pellets were then resuspended and DNA ends ligated followed by a de-crosslinking in the presence of proteinase K and recovering of DNA by isopropanol precipitation. 5 ľg DNA of each 3C library was sheared, purified, and processed into a paired-end library using the Illumina Paired-End DNA sample Prep Kit for sequencing on a NextSeq Illumina platform.