User:David Johnston Monje/Notebook/Maize Endophyte Biofertilizers/2010/05/14

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The Daily Grind

  • Measuring OD600 of O/N of constructs with and without inducer (at 19 hours post innoculation it was Lac::ACC Deaminase - M9 + IPTG/M9 + ACC/M9 + ACC + IPTG = 0.066/0.048/0.093 while Arabad::ACC Deaminase - M9 + Arabinose/M9 + ACC/M9 + ACC + Arabinose = 0.043/0.064/0.049). In SOC without or with Arabinose, ODs of Ara::GFP, Ara::alsSD, Ara::PDS RNAi, and Ara::TBI RNAi were 0.98/0.79, 1.06/1.29, 1.0/0.95, 0.91/0.89. Re-doing over 48 hours on the weekend using the same conditions, but starting with only 25 ul of OD=1 innoculate per 2 mL tube.
  • Also doing an acetoin production assay with the alsSD construct (and it worked!!! but weirdly, non-induced Arabad::alsSD made more acetoin than induced). Gonna plate these out on three plates of LB, without Arabinose, with 0.01 M Arabinose, and with 0.1 M Arabinose.
  • Want to make PCNM with 0.1 M Arabinose and IPTG (0.5 ul per mL) to test construct filled E. coli effect on potato growth. Gonna dip plants in OD600 = 1.0 cells (grown with antibiotics, washed and resuspended in phosphate buffer) and do it in triplicate.