Figure 3.

RB-TnSeq analysis of biofilm forming cultures of the pooled TnSeq library. (A) Diagram of the RB-TnSeq library biofilm formation experiments and collection of samples. Triplicate samples were pooled prior to DNA extraction to ensure enough biomass and material for barcode sequencing. (B) Representative biofilms, or lack thereof, left attached to the glass after decanting and washing steps for the RB-TnSeq library, WT, a homogeneous biofilm formation strain, and a 1:1 mixture of WT and the biofilmation strain. Volcano plots of experiment 1 (E1) test tube (C) biofilm (BF) fractions and (D) settled fractions (S), and experiment 2 (E2) test tube fractions in BG-11 (E) biofilm, (F) settler, and (G) planktonic (P) fractions, presented as described for Figures 1B,C except that the x-axis is the fitness value and the y-axis is the T-value statistic. (H) Comparison of hit data for ORFs with significant fitness value changes, presented as described for Figure 1D, for all BG-11 test tube fractions. (I) Enrichment analyses for pili and known biofilm genes in the clusters of hits identified in (H) are presented as described for Figure 1E.