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

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.