title: GSM5737619: Adh excess B; Neisseria gonorrhoeae; RNA-Seq
library_construction_protocol: Suspension: Bacteria from 16 wells were pooled together in a 15mL conical tube, and two volumes of RNAprotect bacteria reagent (Qiagen 76506) were added. Adherent: supernatant was removed and 1 volume (500 μL) KSFM-GF and 2 volumes (1 mL) of RNAprotect bacteria reagent (Qiagen 76506) were added. N. gonorrhoeae was collected by scraping, and bacteria from 8 wells were pooled together in a 15mL conical tube. Both: bacteria were incubated at room temperature for 5 min, and pelleted at 4,000 x g for 11 min.Samples were lysed according to the “Enzymatic Lysis and Proteinase K Digestion of Bacteria” protocol from Qiagen before proceeding with RNeasy Plus mini kit (Qiagen 74134) per manufacturer's instructions. Ribosomal RNA reduction was completed using NEBNext rRNA Depletion Kit (Bacteria) (New England Biolabs E7850), and RNA library prep was done using NEBNext Ultra II Directional RNA Library Prep Kit (NEB E7760).