GSM1215227: WT2; Neisseria gonorrhoeae; RNA-Seq
Total RNA was isolated from RNAprotect-treated pellets using chicken egg white lysozyme (Sigma) and the RNeasy kit (Qiagen).  Turbo DNA-free enzyme (Applied Biosciences) was used to deplete total RNA of any contaminating DNA.  We further purified our samples by magnetically depleting ribosomal RNA two times using the MICROBExpress mRNA enrichment kit (Ambion).  Global cDNA was made using random hexameric primers (Invitrogen).  Agilent Bioanalyzer analysis was performed following DNA removal, mRNA enrichment, and cDNA library production to ensure sample quality.  The double-stranded, non-directional cDNA libraries representative of the transcriptomes of all three WT and all three misR::kan mutant cultures (see Passalacqua et al., 2009; Croucher and Thomson, 2010) were sent to the Emory GRA Genomics Core for library amplification and sequencing on the Hi-Seq2000 next generation sequencing platform (Illumina).
GEO
GSM1215227