Team:HokkaidoU Japan/Notebook/aggregation Week 13

From 2012.igem.org

September 26th

Characterization of Aggregation

Characterization of Aggregation by pBAD-RBS-eCFP-RBS-Ag43-dT-pSB1C3 construct.

  1. Prepared 5 ml LB liquid medium with appropriate antibiotics into round bottom glass tubes that capped with sponge like cap.
  2. Suspended colonies into the medium.
  3. Incubated at 37C for 12 hrs at 180 rpm. For this incubation, pipetted 150 ul of this cultivating medium to 96-well microtiter plate every 2 hrs.
  4. Analyzed the Absorbance and fluorescence intensity by microtiter plate reader and fluorescence imager.

Result
We couldn't observe eCFP expression, but OD600 of suspension has difference between Arabinose added culture and negative control. Data(OD600)

2(hour) 4 6 8 10 12
Ara+
0.0119457 0.1101659 0.013273 0.1486576 0.2707692 0.3663348
Ara-
0.0119457 0.1499849 0.2813876 1.0419305 1.1932427 1.221116