Team:Aalto-Helsinki/InterLab

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InterLab 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.

Results

Sample	J23117 + I13504	J23101 + I13504	J23151	Empty backbone	Without backbone	Blank
1	D1	D2	C1	C2	C3	B
1	D1	D2	C1	C2	C3	B
1	D1	D2	C1	C2	C3	B