Team:NAWI Graz/InterLab

INTERLAB MEASUREMENT STUDY


Reliable and repeatable measurement is key to compare and analyse data from different labs all over the world. To support the effort to create detailed protocols to measure fluorescence and improve the possibility of comparing data, the iGEM Team NAWI_Graz 2017 decided to participate in the [Fourth International InterLaboratory Measurement Study] in synthetic biology. The goal is to establish a GFP measurement protocol based on engineering principles.

Out tasks were to follow exactly the [iGEM protocol], fill in the prepared excel sheets and send the results to the iGEM headquarter.


Results

For this study we needed to transform 8 devices into the presupposed bacteria strain E.coli DH5-alpha. This included a positive Control [I20270], the constitutive TetR repressible promoter as negativ Control [R0040], Test Device 1 [J364000], Test Device 2 [J364001], Test Device 3 [J364002], Test Device 4 [J364003], Test Device 5 [J364004] and Test Device 6 [J364005]. They are all stored in pSB1C3 and were cultured on LB-Agar plates with a chloramphenicol concentration of 100 µg/ml. As we had grown colonies the day after the transformations, we could start with the cell measurement protocols.

Calibration Protocols

OD600 Reference Point

Results of OD600 measurement of 1 ml LUDOX and 1 ml H²O.
[protocols table]

Protocol Fluorescein Fluorescence Standard Curve

We measured the in PBS diluted fluorescein 96 well plate in a plate reader with the settings XY and turned of pathlength correction.
[flourescence table]

Cell Measurement Protocol

  • Day 1 OD600 Measurement
  • Day 2 Transformation
  • Day 3 Overnight culture

InterLab

Bronze Medal Criterion #4

Standard Tracks: Participate in the Interlab Measurement Study and/or improve the characterization of an existing BioBrick Part or Device and enter this information on that part's Main Page in the Registry. The part that you are characterizing must NOT be from a 2017 part number range.

For teams participating in the InterLab study, all work must be shown on this page.