User:David Johnston Monje/Notebook/Maize Endophyte Biofertilizers/2009/07/09

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The Daily Grind

  • Am going to 96 well plate out bugs on Kan R2A, into 96 well VP media, and am going to plate E. coli on M9 media (without nitrogen source) with 0.3 g/L of filter sterilized ACC, but in the future would like to try out the following assay:

DF-ACC salts minimal medium was used to check if bacterial strains had ACC deaminase activity. DF salts minimal medium (Dworkin and Foster 1958) contained the following: KH2PO4, 4.0 g; Na2HPO4, 6.0 g; MgSO4·7H2O, 0.2 g; FeSO4·7H2O, 1.0 mg; H3BO3, 10 μ g; MnSO4, 10 μ g; ZnSO4, 70 μ g; CuSO4, 50 μ g; MoO3, 10 μ g; glucose, 2.0 g; gluconic acid, 2.0 g; citric acid, 2.0 g; distilled water, 1 L. DF-ACC salts minimal medium was supplemented with 0.3033 g of ACC in each liter of DF salts mini- mal medium as a sole N source. Solid media were obtained by the addition of 12 g of water-washed agar to these salt solutions. The heat-labile ACC was filter-sterilized through a 0.2-μ m membrane, and the filtrate was added to the DF salts minimal medium after autoclaving.

  • Nafisa cut up a bunch of potatoes for me.