Introduction

The 2016 UCLA iGEM Team is proud to participate in the Third International InterLab Measurement Study in synthetic biology. As members of the synthetic biology community, we are committed to providing robust data for the development of novel characterization methods in the rapidly growing biological design fields of synthetic biology. The purpose of the 2016 InterLab study is to examine the precision and accuracy of fluorescence data and information processing from active iGEM teams participating around the world. By collecting fluorescence data from multiple teams in absolute units, variability in measurement and consistency of data collected from instrumentation following a uniform procedure can be determined within a high degree of accuracy. This notebook will record all protocols, daily experiments, relevant images, as well as the raw data and graphs used in the Interlab Submission Form.

Methodology

• The main protocol applied for the 2016 Interlab Study was provided by iGEM Headquarters, and can be found here.
• The specific transformation protocol used was for New England Biolabs C2989K (E. coli K12 strain) and can be found here

Results

The following results are the fluorescence calibration curve (FITC) and the cell sampling OD600 and fluorescence results. The graphs were generated from the Excel spreadsheet provided by iGEM.

FITC Fluorescence Calibration Curve

Cultures were sampled at the t=0, 1, 2, 3, 4, 5, and 6 hour marks in 100uL aliquots from 5mL cultures and stored on ice to arrest growth until ready for measurement. These results are almost as expected, there was highly stagnated growth for both replicates of Device 1; we attribute this discrepancy to human/experimental error.

OD600 Graph for the Positive & Negative Control and Devices 1, 2, and 3

Following the last sampling at t=6hr, all samples were added to a 96-well plate in preparation for fluorescent measurement. The plate layout can be found in the lab notebook below. Fluorescent values were normalized following measurement,

Fluorescence Graph for the Positive & Negative Control and Devices 1, 2, and 3

Notebook