Journal: bioRxiv

Article Title: Design of a new model yeast consortium for ecological studies of enological fermentation

doi: 10.1101/2024.05.06.592697

Figure Lengend Snippet: Cytometry set up with an example of sample at inoculation. A: YL2 (600DLP-620/15)/BL1 (495DLP-530/30) dot plot is used to separate GFP positive and mCherry positive cells. B: VL2 (495DLP-512/25)/SSC dot plot derived from the GFPPos gate on A, is used to separate mCitrine (-) from eGFP signal (+). C: VL1 (417LP-440/50)/SSC dot plot derived from the GFPNeg gate on A, is used to separate non-fluorescent cells from BFP positive cells. D: Hierarchy of gating and percentage of each population. % Total indicates the % of the total population of events in the corresponding gate.

Article Snippet: The following plasmids were constructed by Gibson assembly (Gibson et al., 2009): pFA6a-link-yEmCitrine-NATMX, pFA6a-link-yomCherry-NATMX, pFA6-TDH3.1kb.Td-mCitrine-NATMX, pFA6-Hu1kb-BFP2-KAN. Gibson assembly were done using the NEB Builder HiFi DNA Assembly Master Mix (New England Biolabs) and transformed into E. coli DH5ɑ (New England Biolabs) following the manufacturer instructions. pFA6 plasmid backbone, antibiotic resistance cassettes ( KanMX, hphMX ) and fluorescent protein genes (EGFP, mCherry, mCitrine, mTagBFP2) were obtained from plasmids ordered from AddGene (#44900, #44899, #44645, #44903, Supplementary Table 1 , ; ).

Techniques: Cytometry, Derivative Assay