InterLab Study

Our InterLab Study Lab Book can be found here for a fully detailed overview of our study.

Devices

The following devices were created in a backbone pSB1C3:

⚫ J23101 + I13504 (B0034-E0040-B0015)
⚫ J23106 + I13504 (B0034-E0040-B0015).

However, constructing a device J23117 + I13504 (B0034-E0040-B0015) wasn't successful due to the time limit and limited ligation times. More details available on Lab Book. The device sizes were analyzed with restriction mapping which results can be seen in Figure 1.

Used BBa_I20270, a GFP expressing part in the pSB1C3 backbone as a positive control. Negative controls were organisms with an empty plasmid and without any vector.

Protocols

Constructing devices

Protocols for the restriction, ligation and transformation of BioBricks can be found here.

Cultivations and measurement

Followed the next protocol for preparing the samples:

Streaked out LB-plates with E.coli TOP10 strain containing every device and control with chloramphenicol concentration of 35ug/ml. Incubated plates overnight (18-20 hours) at 37C. Picked up biological triplates from the plates and inoculated 3 ml liquid cultures from the colonies with 12ml polypropylene test tube. Incubated the tubes at 37C with shaking at 300rpm for 16-18 hours. Measured OD600 values of cultures in cuvettes and calculated the dilution required for each sample to obtain the OD value of 0,5. Diluted and re-measured the samples to reach 5% accuracy of 0,5. Transfered 200ul of each sample into 96-well transparent plate and set the platereader to read fluorescence intensity. Proceed with the measurements.

Results

1st Technical Replicon

2nd Technical Replicon

3rd Technical Replicon