Phages weaponize their bacteria with biosynthetic gene clusters
Autor: | Paul J. Kempen, Ákos T. Kovács, Anna Dragoš, Carlos N. Lozano-Andrade, Mikael Lenz Strube, Aaron John Christian Andersen |
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Rok vydání: | 2021 |
Předmět: | |
DOI: | 10.5281/zenodo.4449745 |
Popis: | Method of data acquisition: Overnight cultures of selected strains were obtained, optical density was measured and cultures were pelleted down (8000 g, 5 min) and re-suspended in 0.9 % NaCl, to reach optical density of 5. Next, 1:1 co-cultures were created by mixing equal volumes of selected strains suspensions. Such co-cultures were then inoculated at 1 % into 200 µl of LB distributed in 96-well microtiter plates. Cultivation was performed in Synergy XHT multi-mode reader (Biotek Instruments, Winooski, VT, US), at 37 ºC with linear continuous 508 shaking (3 mm), monitoring the optical density (600 nm) as well as GFP (Ex: 482/20; Em:528/20; Gain: 35) 509 and mKate (Ex: 590/20; Em: 635/32; Gain: 35) fluorescence every 10 min. For long-term competition assay, co-cultures were transferred to fresh LB medium (2.5 % inoculum), every 24 h. Usage Notes: Raw data from all time-lapse experiments using Plate reader, were complied into single excel file. Data from separate experiments are presetned on different excel sheets. In the upper left corder, some default information on the experimetnal setup are provided (Synergy XHT multi-mode reader automatically provides these data during export). Raw data are arranged in columns: I column shows time interval, II column shows temperature, and the next columns shown number of well in 96-well plate (from A1 to H12). Above the A1-H12 columns, explanatory text was added to identify which culture (or co-culture) grows in each well. As strain names are pretty long, these explanatory text was coded: letters for monoculures, and comp.+letter for co-cultures (comp=competition), a legend above raw data was provided to explain all labels. Legend also connects raw datasets with figures in the mansucript. On the right, average from biological replicates (laballed: repliate 1, replicate 2...is calculated for each timepoint). Each sheet contains optical density data, GFP fluorescence data and mKate fluorescence data (starting from top to bottom) Work supported by Danish National Research Foundation (DNRF137) for the Center for Microbial Secondary 449 Metabolites |
Databáze: | OpenAIRE |
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