Team:USNA Annapolis/InterLab


Experimenting With DNA


History and Purpose

These past three years, iGEM has created two of the biggest interlaboratory studies ever done in synthetic biology. These studies establish a baseline of reciprocity. The purpose of these studies is to maintain laboratory standards over all of the iGEM groups. This ensures the participating groups have a baseline competence in synthetic biology.

Synopsis of Procedure

The procedure for the Interlab study was broken into three components with each having a different purpose. The goal of the first part of the study to calculate a numerical conversion factor to convert an absorbance reading taken from the plate reader to an OD600 reading. This was accomplished by using LUDOX-S40 as a reference point and subtracting the absorbance from the water. The second section of the protocol was the creation of a fluorescein standard curve which was done by resuspending fluorescein in 1x PBS then making a serial dilution from 100% fluorescein to 100% PBS. The fluorescein concentrations can then be converted into a concentration of GFP. The final part of the study involved transforming the provided test devices, inoculating cultures and growing them in the incubator, then measuring the absorbance and fluorescence at time 0, 2, 4 , and 6 hours. These readings were then converted into GFP concentrations using the fluorescein standard curve.