GSM9340360: CDM with iron, biol rep1; Neisseria gonorrhoeae; RNA-Seq
Bacteria from 3 replicate wells (12 mL total) were pooled together in a 15-mL conical tube and pelleted at 4,000 x g for 11 minutes. Gc was resuspended in 200 μL RNAprotect bacteria reagent (Qiagen 76506) and stored at -80 °C until processed to purify RNA. Samples were lysed according to the “Enzymatic Lysis and Proteinase K Digestion of Bacteria” protocol from Qiagen before proceeding with RNeasy Plus minikit (Qiagen 74134) per manufacturer's instructions. DNase treatment was completed using a TURBO DNAfree kit (Invitrogen AM1907). rRNA reduction was completed using NEBNext rRNA Depletion Kit (Bacteria) (NEB E7850). Strand specific RNA libraries were prepared for sequencing using standard Illumina protocols using NEBNext Ultra II Directional RNA Library Prep Kit (NEB E7760).