Calibrated fluorescent materials
A significant challenge in the field of synthetic biology is the ability to obtain consistent and reproducible measurements across different labs. iGEM's International Interlaboratory (Interlab) Measurement Study has played an important role in tackling this challenge1. This year, our team has chosen to participate in the seventh edition of the Interlab study! In this edition of the study, the aim has been extended to include measurement procedures from one-color calibrations to multi-color calibrations. For more details, please visit the iGEM technology page .
The aim of the calibration experiment was to set a baseline with our equipment by measuring known quantities of materials. We measured dilutions of a known concentration of four components: Fluorescein Sodium Salt - green fluorescent,
Sulforhodamine-101 - red fluorescent, Cascade Blue - blue fluorescent, and Monodisperse Silica Nanoparticles - optical density(OD)/bacterial cell count calibrant. We used a
Tecan Plate reader Infinite 200 PRO with a
clear bottom black 96 well plate
for this and all subsequent measurements.The submitted results can be found below:
Submitted results
The aim was to compare an array of six fluorescent protein devices against the calibration previously carried out. The goal of this was to gauge how single fluorescent protein expression or independent double protein production varied between labs. This included six plasmids with red, blue, or green fluorescent proteins, or a combination of two under independent transcriptional units. We transformed these into DH5α E.coli and normalized their culture density in accordance with the protocol, and measured in a 96 well plate.
Due to some transformation troubles, all plasmids first had to be transformed to TOP10 E.coli before the plasmids were harvested and transformed into DH5α.
The submitted results can be found below:
Submitted results
The aim of this experiment was to answer a fundamental question: does the order of transcription units influence which one is expressed the most strongly? Six plasmids with combinations of two the Red, Blue, and Green fluorescent protein genes under the same promoter were transformed into DH5α. Their culture density was normalized in accordance with the protocol and measured in a 96 well plate.
Due to some transformation troubles, all plasmids first had to be transformed to TOP10 E.coli before the plasmids were harvested and transformed into DH5α.
The submitted results can be found below:
Submitted results
This experiment aimed to measure the interlab consistency of plate culturing vs tube culturing on dynamic green fluorescence and OD. We used six devices in DH5α E.coli which all coded forms of green fluorescent proteins, diluted to specification, and carried out measurements in a 96 well plate.
This line was carried out first as we were able to use aliquoted transformants that had already been transformed by previous iGEM teams to save time.
The submitted results can be found below:
Submitted results
The raw data for each section can be found below:
Raw data