Team:MIT/InterLab

• BACKGROUND
• EXPERIMENTS
  Synthetic Promoters microRNA Recombinase Proof of Concept Collaborations
• FUTURE WORK
  Circuit Design Clinical Applications
• HUMAN PRACTICES
  Parts Toolbox Accessible Protocols Integrated Human Practices Public Outreach Interlab Study
• NOTEBOOK
  Lab Notebook Parts Mammalian Parts Collection Attributions Safety
• AWARDS
  HP Silver HP Gold Integrated HP Basic Part
• TEAM

Human Practices

iGEM Headquarters is conducting a study to explore the question, "How close can the numbers be when fluorescence is measured all around the world?"

MIT iGEM participated in this study by measuring GFP fluorescence in the samples provided by iGEM HQ and reporting the results.

E.Coli Flow Cytometry Protocol (from ... via Nicholas DeLateur)

Day 0:
A single isogenic colony is picked up with a sterile pipette tip and used to inoculate a well in a 96-well plate containing 200 uL of M9-Glyc-Kan media. The 96-well plate is shaken at 900 RPM at 37 C for 12-16 hours overnight.

Day 1:
Each well is diluted twice, 15 uL into 185 uL of M9-Glyc-Kan.
The subculture is grown at 900 RPM at 37 C for 3 hours.
The subculture is diluted 5 uL into 145 uL of M9-Glyc-Kan and grown for 5 hours at 900 RPM, 37 C
Transfer 5 uL of the subculture to 195 uL of 1 X PBS with 2 mg/mL Kanamycin.
Allow to sit on bench for 1 hour.
Run flow Cytometry.