Team:The Tech Museum/Notebook

Notebook

Timeline of major events:

• May: Basic concept research and design
  
• June: Software development initiated; plasmid design started and promoter-RBS’s picked
  
• July: Wetlab testing of possible color protein combinations
  
• August 10: Design of complete tri-color plasmids finalized
  
• August 24: Assembly of tri-color plasmid pools completed (DNA2.0)
  
• September 10: Tri-color plasmids optimized on museum wetlab setup
  
• September 29: Software fully integrated with hardware on mobile exhibit
  
• September 30 - October 3: Prototyping of mobile exhibit with museum staff and visitors
  
• October 6 - October 17: Data collection on the museum floor with visitor!

Biology Details:

Design of tri-color plasmid pool (June 2014)
9 promoter-rbs pairs of varying strength from Kosuri et al paper (2013):

Selection of colored reporter proteins
Co-transformation testing of multiple color combinations of proteins from DNA2.0
Transformations on Kanamycin and IPTG

Final tri-color plasmid designs, assembled by DNA2.0
Chromogenic plasmid pool
Blitzen (blue)
Kringle (yellow)
Paprika (red)
Fluorescent plasmid pool
CindyLouCFP (400/495)
KringleYFP (520/542)
PaprikaRFP (554/590)

Optimization of plasmid pools with visitor-accessible museum wetlab set up
Conditions
6cm plates
400 ug/ml Amp
0.3 ul unamplified plasmid pool in 100ul CaCl2
20 ul competent bacteria
Current visitor wetlab transformation protocol:
30 sec on ice
40 sec heat shot at 42 degrees C
Maturation times
Chromogenic pools ~ 5 days 37 degrees C
Fluoroescent pools ~ 3 days 37 degrees C
Low copy fluorescent plasmid pool gave more reliable results with most color variety

Safety:
We did not use any dangerous organisms or reagents. Transformation of bacteria was done in the musuem’s existing wetlab setup.