User:David Johnston Monje/Notebook/Maize Endophyte Biofertilizers/2010/04/08

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The Daily Grind

  • Met Manish and discussed second rep of growth promotion results. Agreed we have to understand ideal conditions for potato bioassay - will repeat with three isolates Burkholderia phytofirmans (2A10), Dionococcus (2D3), and Methylobacterium (2D10). Will treat potatoes either with 1 mm looped colony resuspended in 1 ml of sodium phosphate buffer or R2A (measure resulting OD - control to 0.1). Will also make O/N in R2A and phosphate buffer (resuspend colony to OD1 if not enough growth) control OD so that they're 1.0 (as opposed to OD for suspension which should be about 0.1)
  • Dipping plants in O/Ns and immediately sampling some for microbial counts. Plantlets will be put into tubes momentarily, re-isolated (the ones for day 0 sampling) and crushed in eppendorfs. 100 ul of LB will be added for dilution, and 1 ul of this will be resuspended into 99 ul of LB. This will be further diluted 1:99 in LB. 50 ul of these later two dilutions will then be spread on half plates of TSA + chloramphenicol with Xgal and IPTG on them.