Transcriptomic analysis of C. jejuni grown in anaerobic jar with air mix and in Modular Atmospheric Controlled System
ABSTRACT: Transcriptomic analysis of C. jejuni HPC5 grown in anaerobic jar supplemented with air mix and in Modular Atmospheric Controlled System (MAC). C. jejuni HPC5 is a Campylobacter strain isolated from broiler chickens. Majority of the Campylobacter strains are cultured in an atmosphere containing 5-10% (v/v) oxygen or 5% hydrogen and 5-10% (v/v) carbon dioxide and the rest with nitrogen. To set up a jar (3.5L, Oxoid) to the required microaerophilic conditions, a vacuum is created to -22 psi and then air mixture of 85% v/v nitrogen, 10% v/v carbon dioxide and 5% v/v hydrogen (Air Products, Crewe, UK) is introduced into the jar. This resulted in a microaerobic atmosphere containing approximately 5.6% oxygen, 3.6% hydrogen, 7.3% carbon dioxide and 83% nitrogen. The Modular Atmospheric Controlled System (MAC; Don Whitely Scientific) is an anaerobic chamber where air mixture of 85% nitrogen, 10% carbon dioxide and 5% oxygen is used for growing Campylobacter. Differences in growth pattern were observed when Campylobacters were grown in it and transcriptomic analysis was done to prove the difference in the regulation of genes in the two conditions. Overall design: There are three biological replicates for each experiment. Seven independent cDNA preps of C. jejuni HPC5 were prepared and they were labelled independently using AF555 (Fisher, UK). The labelled cDNAs were mixed together and this acted as the control. C. jejuni HPC5 was grown both in the Jar and in the MAC and their cDNAs were labelled independently with AF645 (Fisher, UK). Hybridisations were done between C. jejuni HPC5 grown in jar and in MAC in triplicates. The supplementary file (linked at the foot of this record) represents the averaged normalised values for each experimental condition (3replicates/experimental condition).