Difference between revisions of "Team:Lethbridge/InterLab"

Overview

The ability for parts to provide repeatable outputs is key to engineering applications, including synthetic biology. The purpose of the 2017 interlab study was to address how reproducible and at what levels protein expression could be driven by standardized promoters and ribosomal binding sites. Constructs were provided by the iGEM measurement committee that utilized green fluorescent protein as a reporter. This allowed interlab study contributors to easily follow protein expression driven by the standardized promoters and ribosomal binding sites.

In order to better compare expression levels between research groups, the 2017 interlab study incorporated calibration protocols to allow for reporting of fluorescence levels in absolute levels. Specifically, fluorescence levels from biological samples were calibrated against fluorescence from a fluorescein standard curve, allowing for fluorescence results to be reported in terms of the amount of fluorescence produced by a certain concentration of fluorescein.

Test Devices

The negative control for the interlab study did not contain DNA coding for recombinant fluorescent-protein expression , thus was well-suited for examining background fluorescence of cellular components. The remaining constructs contained GFP (E0040) under the control of various promoters and ribosomal binding sites. The promoters and ribosomal binding sites utilized for Test Devices 1-6 are outlined below. All constructs contained E0040, B0010, and B0012 following the promoter and ribosomal binding site.

Protocols

Methods

Results