Strain MK350 was grown under the same conditions as used when growing cells for microscopy analysis; cells of OD600 = 0.4 were diluted 100-fold in C+Y medium with 0.1 mM ZnCl2 and incubated for 2.5 hours until OD600 = 0.15. Cells were then harvested by centrifugation for 5 min at 6500 x g at 4°C. Genomic DNA was isolated using the Wizard® Genomic DNA Purification Kit (Promega) as described previously (Slager et al. 2014 Cell). Fragmentation was performed using Covaris instrument, and libraries were prepared using NEBNext Ultra DNA Library Kit for Illumina prior to sequencing on an Illuminia HiSeq 2000 Platform (all performed at the Genomics Core Facility, EMBL, Heidelberg, Germany).
Views: 2167 Downloads: 108
Created: 19th Jul 2016 at 09:40
Last used: 21st May 2019 at 16:33