User:David Johnston Monje/Notebook/Maize Endophyte Biofertilizers/2013/07/16

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Remains of the Day

  • Got back from vacation today and Stephanie is gone so I'm gonna try orient myself on my own
    • Find photos from tomato disease assay and evaluate (see below - looks like DC95-5 gives the most visible lesions - will design experiment and plant more tommorow)
    • Look through folder for clone sequences and colony PCR sequences that Athir sent in (the best growth promoting potato endophyte was Methylobacterium fujisawaense strain 7RN-5A (EU379312), the large 408 bp ITS4 BstNI cut peak in molasses rhizosphere samples was an uncultured Galactomyces, and large 668 bp ITS4 BstNI cut peak in molasses roots samples was Olpidium brassicaceae)
    • Went over some PCR results with Steph once she came back and found that the nested Pythium irregulare primers worked well, but the Pythium ultimum, Phytopthera fragariae, and Phytopthera infestans primers might need a bit of optimizing (will work with her on Thursday to do this). She also did the 27F Degen/1492Rh primer testing, and had troubles, so I'll do a test tonight on a couple different soil samples (E. coli, Sick Arrugula Rhizosphere, Trial 3 Water Treated Strawberry Rhizo 1, 2 and 3, and Trial 3 Pesticide and Bacillus/Pseudomonas/Burkholderia Soaked Strawberry Rhizo 1, 2 and 3) and at a couple different annealing temperatures (48, 50, 52, and 54), with 40 cycles of PCR. In 25 ul of reaction each.