Team:WPI Worcester/InterLab


Interlab

As part of our laboratory work, we decided to participate in the 4th annual interlaboratory (interlab) study conducted by the iGEM foundation. The interlab study aims to address the problem of reproducibility in the field of synthetic biology. By providing a kit of parts, a comprehensive protocol, and a data analysis spreadsheet to all teams who participated in the study in order to see whether this would minimize error associated with reproducing experiments. Teams were asked to transform six GFP expressing constitutive devices into DH5α E. coli cells and then measure the fluorescence and absorbance of each device. The fluorescence and absorbance of a positive and a negative control was also measured. The devices used and their associated iGEM parts page are listed below:

Positive Control (BBa_I20270)

Negative Control (BBa_R0040)

Test Device 1 (BBa_J364000)

Test Device 2 (BBa_J364001)

Test Device 3 (BBa_J364002)

Test Device 4 (BBa_J364003)

Test Device 5 (BBa_J364004)

Test Device 6 (BBa_J364005)

Given the fact that our team had a plate reader easily accessible to us, we chose to obtain our measurements using a plate reader as opposed to using a flow cytometer. Before measuring the fluorescence and absorbance of each device, we created standard curves for each. Both standards were made using the solutions provided by the iGEM foundation in the Measurement Kit. The protocol for creating the standards, and for obtaining the measurements for our devices can be found here.

We were able to create a standard curve for fluorescence using the following data, and our raw data can be found here: InterLab Table 1

The associated figures can be found below:

InterLab Graph

Figure 1





InterLab Graph 2

Figure 2



We were able to create a standard curve for absorbance using the following data:

InterLab Table 2



After developing standards for both fluorescence and absorbance, we measured the fluorescence and absorbance for two colonies for each device.

Average Raw Fluorescence:

InterLab Table 3

Average Raw Absorbance:

InterLab Table 4


Flueorscent to OD Ratio for Colony 1:

Colony 1 Chart

Figure 3


Flueorscent to OD Ratio for Colony 2:

Colony 2 Chart

Figure 4


Average Flueorscent to OD Ratio:

Colony 1 and 2 Averages Chart

Figure 5