Simple Growth curve on M9 |
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In our quest for Electro-genetic responsive promoters, we used the E. coli Promoter Collection, a library of close to 2000 E. coli promoter fused to a Green Fluorescent Protein (GFP) [1]. To determine experimental parameters for following experiments, some basic information about the behaviour of these strains are necessary. |
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The present experiment aims at looking at the growth of the strain use in further experiements |
Materials and methods |
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The E. coli Promoter collection is delivered in 21 separate 96 well plate. Each well hold a glycerol stock of a specific promoter-GFP of the collection. As all of most of experiments will be conducted through pooling individual plates, it made sense to look at growth curves of each plate separately. Here, we pooled each of the first 5 plates (from AZ_01 to AZ_05, see full detail of the promoter collection for more information) in single tubes. We then measured the growth on a plate reader of each of the 5 pooled population, in triplicate. |
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We grew the cells to saturation overnight in M9 Media (0.4% Glucose, Amino Acid Complete). In the morning, cultures were re-inoculated in fresh M9 (to a ratio 1:100) and grown in a Tecan Infinite® 200 PRO for 19 hours to measure OD600. |
Results |
We observe that all population reach different final OD. This might imply different burden for different plasmids
Future Work |
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References |