Mycoplasma feriruminatoris transcriptome

The goal of this experiment is to determine the overall relative strength of promoter sequences in Mycoplasma feriruminatoris. For this, 2 replicates were grown in parallel in Hayflick media and the RNA wa extracted at exponential growth phase (20 hours). With this data, new promoter sequences could be designed and further validated by the use of RT-qPCR and reporter assays. Overall design: The cultures were started at 1:1000 proportion (10 µl in 10 ml of growth media). They were left to grow for 20 hours and collected at exponential phase. The cells were separated and washed via centrifugation. The cellular pellet was then extracted using a standard RNA extraction method using the RNeasy kit from Qiagen. The stage of exponential growth phase was confirmed via CFU/ml counting.