Figure 1. Liquid triplicate cultures of each construct
1. The first device is [http://parts.igem.org/Part:BBa_I20260 BBa_I20260], which is a biobrick part that contains:
A promoter, [http://parts.igem.org/Part:BBa_J23101 BBa_J23101], a downstream gene ([http://parts.igem.org/Part:BBa_E0240 BBa_E0240]), and a backbone ([http://parts.igem.org/Part:pSB1C3 pSB1C3]).
The downstream gene is also a composite of a number of biobrick parts, including an RBS - [http://parts.igem.org/Part:BBa_B0032 B0032], a GFP coding sequence - [http://parts.igem.org/Part:BBa_E0040 E0040], and a transcriptional terminator - [http://parts.igem.org/Part:BBa_B0015 B0015].
2. The second device was cloned in lab, as described in the interlab study procedure.
The device is composed of [http://parts.igem.org/Part:BBa_J23101 BBa_J23101] and [http://parts.igem.org/Part:BBa_E0240 BBa_E0240], which are the same parts as in the first.
However, the backbone of this device is different, [http://parts.igem.org/Part:pSB3K3 pSB3K3]
3. The third device was also cloned in lab, as described in the interlab study procedure.
The device is composed of [http://parts.igem.org/Part:BBa_J23115 BBa_J23115] and [http://parts.igem.org/Part:BBa_E0240 BBa_E0240], which is the same downstream gene as in devices 1 and 2. However, the promoter sequence of device 3 is different.
This device is in [http://parts.igem.org/Part:pSB1C3 pSB1C3].
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
Microplate Reader Data
Figure 2. Fluorescence Data for the three devices.
Data was collected using [blah plate reader and a 96 well plate blah] with ?50 ul of culture? per well. As can be seen in Figure xx, ...
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