title: Non-adherent transposon mutant library after second infection of a fibrin matrix under flow conditions, first biological replicate
design: A fibrin matrix was formed on an Ibidi -Slide I Luer 3D by incubating 10 mg/mL of plasminogen-depleted human fibrinogen  in PBS with 1.0 KU/mL thrombin in each well at 37C overnight. A colony of the S. canis mutant library after the first selection round was grown overnight in THY (THB + 1.5% yeast extract). The culture was washed once with PBS and resuspended to OD600 of 0.05 in Brain-Heart-Infusion-Broth (Roth) with 2% (w/v) glucose (BHIgluc) added. This suspension was added to the fibrin matrix and the Ibidi system was filled with BHIgluc. Biofilms were allowed to grow for 24 hours under physiological shear stress conditions (10 dyn/cm2) before harvesting non-adherent bacteria by washing the matrix with PBS. The non-adherent bacteria were grown overnight at 37C and genomic DNA was extracted.