GSM8146053: Late_NoMo_rep_A; Methanococcus maripaludis; RNA-Seq
Cultures of MmS2 were harvested during late log phase (24 hrs. for molybdate and molybdenite conditions, 30 hrs. for the no Mo condition) of growth for RNA extraction. Cultures were brought into an anaerobic chamber and were vacuum-filtered over 0.2 µm Supor membrane filters that were then transferred into 2 mL microcentrifuge tubes. The tubes and their contents were immediately flash frozen in liquid nitrogen and stored at -80 °C. Total RNA was extracted from the filters using the Trizol method. DNA contamination was checked by performing PCR amplification using archaeal-specific 16S rRNA primers. rRNA depletion was conducted using custom Methanococcus-specific oligonucleotides, and sequenced on the Illumina NovaSeq platform with 2x150 bp paired-end reads at the University of Wisconsin sequencing center.