Team:Brown/Notebook Meetings/8-3-09

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iGEM Meeting 8-3-09

SFH 218

  • 9:06: UTRA poster presentation. Call Kinko’s and let them know. Or contact Judy Nathanson for printing next door for much cheaper.
  • 9:08: Gary will be out Thursday to Thursday. Poster practice on Wednesday.
  • 9:13: Team 1 update
    • EV131 is in pNoTat
    • Making BL21 competent – need T7 polymerase; greater control over gene expression
    • When running protein gel, run control cell w/o bacteria. To purify protein, will get 6-His antibodies and nickel columns.
    • Possible binding assays – run against stationary histamine. Running free histamine.
    • Threshold level of allergic rhinitis discussed.
    • WE SHOULD INCLUDE ON POSTER the affinity information about EV131
    • Also include considerations.
    • What next after we get EV131 binding? Gary belabors.
  • 9:40: Team 2 update
    • Assembled Sec-GFP. Waiting on primers to amplify and drop into expression vector
    • Also building Trg-EnvZ
    • Working on making S. epi electrocompetent
    • Timeline: make electrocompetent S. epi. Transform with expression vector. By end of week have expression in S. epi.
    • Get a picture of S. epi on the microscope. Screen for contaminants.
    • Also ordering primers for ccdB.
  • 9:46: Gary’s advice about gel extractions
    • Either run multiple lanes and combine, or run a big well
  • 9:55: Team 3 update
    • Now working with RFP as a selection marker
    • Now using assembly service to build OmpC-RBS-RFP-DT. We need to focus on Taz1 mutation.
    • Testing mutagenic primers this week.
  • 10:06: Wednesday at 3PM. Poster practice.