Interlab Study Project

Devices Measured

1. [http://parts.igem.org/Part:BBa_I20260 BBa_I20260] ([http://parts.igem.org/Part:BBa_J23101 BBa_J23101] (strong constitutive promoter) + [http://parts.igem.org/Part:BBa_E0240 BBa_E0240] ([http://parts.igem.org/Part:BBa_B0032 B0032] - RBS, [http://parts.igem.org/Part:BBa_E0040 E0040] - GFP, [http://parts.igem.org/Part:BBa_B0015 B0015] - double terminator))

in [http://parts.igem.org/Part:pSB1C3 pSB1C3] (high copy BioBrick assembly plasmid)

2. [http://parts.igem.org/Part:BBa_J23101 BBa_J23101] (strong constitutive promoter) + [http://parts.igem.org/Part:BBa_E0240 BBa_E0240] ([http://parts.igem.org/Part:BBa_B0032 B0032] - RBS, [http://parts.igem.org/Part:BBa_E0040 E0040] - GFP, [http://parts.igem.org/Part:BBa_B0015 B0015] - double terminator)

in [http://parts.igem.org/Part:pSB3K3 pSB3K3] (low to medium copy BioBrick standard vector)

3. [http://parts.igem.org/Part:BBa_J23115 BBa_J23115] (weak constitutive promoter) + [http://parts.igem.org/Part:BBa_E0240 BBa_E0240] ([http://parts.igem.org/Part:BBa_B0032 B0032] - RBS, [http://parts.igem.org/Part:BBa_E0040 E0040] - GFP, [http://parts.igem.org/Part:BBa_B0015 B0015] - double terminator)

in [http://parts.igem.org/Part:pSB1C3 pSB1C3] (high copy BioBrick assembly plasmid)

Protocols

Sample Preparation

• Inoculate 10 mL of LB with frozen stocks of the three transformed clones (I20260, J23101+E0240, J23115+E0240), a cell background control (TOP10 electrocompetent cells), and an LB only control in a 50 mL Erlenmeyer flask. Grow for 16-18 hours at 37°C, shaking at 300rpm.
• The next morning, add 10µL of each overnight culture to 10 mL of LB with three replicates of each (15 total flasks) and grow 16-18 hours at 37°C, shaking at 300rpm.
• Add 80 µL of each culture to the wells of a clear-bottomed black 96-well plate.

Measurement Protocol

The samples were plated on a 96-well plate as follows:

• The 96-well plate was inserted into the Infinite 200 PRO Microplate Reader
• Open Tecan i-control, the software used to document the readings
• Configure the settings as detailed above
• Obtain fluorescence and OD600 readings

Sequencing Data