User:David Johnston Monje/Notebook/Maize Endophyte Biofertilizers/2010/05/18

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The Daily Grind

  • Did Elena's PCR for her using her protocol. Did two reps of each treatment, including no DNA, Chlamidiya DNA (1 ul per 25 ul tube), and Pi Shoot 3 DNA. Used oil instead of heated lid, and used GoTaq buffer.
  • Screening Arabad::ACC colonies for positives using PCR. With positives, will redo M9 medium + ACC deaminase assay overnight.
  • Remaking soil endophyte plate without contaminated wells.
  • Growing 8 seed endophytes overnight for electroporating them with pDRIVE plasmids tommmorow.
  • Plan to do potato innoculation with E. coli + plasmids. Also planning it with endophytes + plasmids in corn, which will be done alongside Mixteco, B73, and Pioneer innoculated (during seedling stage and watered every week) with Burkholderia phytofirmans taken from Mixteco. Seeds will be germinated on watered tissue paper, dipped in a bacterial suspention of OD600=0.2 and then planted in turface watered with full strength Hoagland's solution. Plants (5 per treatment) will then be grown for one month in greenhouse and watered once a week with 100mL of OD=0.2 Burkholderia.
  • Making overnights of endophytes 1A1, 1A5, 1A8, 1B12, 1C8, 1C12 (no acetoin), 1D9(no acetoin), 1D12, 1E4(no acetoin), and 1G8 to electroporate with Arabad::GFP in pDRIVE.