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Table 1 Outcome of droplet-based microfluidic screening. After FACS sorting of the positive droplets and de-emulsification, cells were plated on solid agar to recover the sorted clones. For each screening condition, the activity toward resorufin-β-GalNAc of 372 of the 22,200 clones recovered on solid plates was quantified at 37 °C in 50 mM Tris/HCl, pH 8 or 9, to select those with at least twice the activity of the E. coli host used as negative control. Then, the clonal redundancy resulting from the bulk de-emulsification of the hit droplets was quantified by sequencing the metagenomic insert extremities for each positive clone, in order to determine the yield of different positive clones isolated from the initial metagenomic library of 20,000 clones

From: Investigating host-microbiome interactions by droplet based microfluidics

  Recovery yielda (%) Positive clonesb (%) Different positive clones Positive clone yieldc (‰)
Library pH 8
 24 h 146 13 14 0.7
 48 h 179 8 15 0.75
 72 h 94 6 9 0.45
Library pH 9
 24 h 47 9 6 0.3
 48 h 100 1 2 0.1
 72 h 28 1 1 0.05
  1. aRecovery yield (%) = 100 × (number of clones recovered on solid plates)/(number of positive droplets)
  2. bPositive clones (%) = 100 × (number of positive clones in quantitative microplate assays)/(number of clones recovered on solid plates)
  3. cPositive clone yield (‰) = 100 × (number of different positive clones)/(initial library size)