User:David Johnston Monje/Notebook/Maize Endophyte Biofertilizers/2009/08/17

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The Daily Grind

  • Grinding 5 three leaf stage plants up (Gaspe) for TRFLP and culturing experiment. They were grown in elora soil, rinsed in sunglight soapy water for 10 minutes, followed by twice in 2% bleach for 10 minutes, a dH2O rince, one 100% EtOH rinse for 10 minutes, and then three rinses in sterile water. Each plant weighed approx 0.3 grams, so adding 300 ul sodium phosphate buffer to each for resuspension and plating/DNA extraction. From the left over in each bowl, I added 300 more ul, took 50 ul from that into 450 ul of fresh buffer, then took 50 ul of that into another 450 ul. Plated 50 ul of that. Also took 50 of each of these initial dilutions, pooled them, and repeated dilution for a pool. Considering doing DNA extraction on these, rather than the plant sample.