title: STAT_LB_MCH1_B
organism: Vibrio cholerae
description: Summary: Wild-type monochromosome (MCH1) variant of V. cholerae grown in LB at 37C with 260 rpm shaking until stationary phase (OD600 of 1.5 or greater). Biological replicate B.Full: Wild-type (EPV50) and wild-type monochromosome (MCH1) variants of V. cholerae strain O1 biovar El Tor str. N16961 were struck on solid agar and grown overnight at 37 C. Three individual colonies were selected from each strain and used to inoculate 10 mL of LB media, which was grown overnight at 37C with 260 rpm shaking. The following morning, these overnight pre-cultures were back-diluted to OD600 of 0.05 in 5 mL of either LB or M9 minimal medium (1X M9 salts, 2 mM MgSO4, 0.1 mM CaCl2, 0.4% glucose, and 10 ug/mL thiamine Hcl). The same biological replicate was used to inoculate both growth conditions. Exponential phase cultures were grown to OD600 of 0.60-0.80. Stationary phase samples were grown to OD600 of 1.50 or greater. To harvest, cultures were centrifuged at 10,000 rpm for 5 mins at 25 C, and the supernatant was removed. gDNA was prepared immediately following harvest using GeneJet Genomic DNA Purification Kit (Thermo Scientific, K0721). Purified gDNA was quantified using Qubit dsDNA HS Assay Kit (Invitrogen, Q32851) and quality was assayed using a Nanodrop 2000 Spectrophotometer (Thermo Scientific, ND-2000). Whole-genome libraries were prepared from purified gDNA using Twist 96-Plex Library Prep Kit (Twist Bioscience, 104950) with dual adapters. Libraries were subsequently pooled and sequenced on Illumina NovaSeq6000 (S4) to a depth of 60,000,000 reads per sample.