Team:Exeter/Interlab
Interlab Study
Chapter 1: New Beginnings
Once upon a time, there was an iGEM team from the University of Exeter. Determined to contribute to the field of synthetic biology, we decided to participate in the Second International Interlab Measurement Study. While determining fluorescence levels for the three Interlab devices, we hoped to gain some experience in the lab before starting our iGEM project, Ribonostics. We aimed to build and measure the fluorescence of three BioBrick devices: 1.J23101 + I13504 2.J23106 + I13504 3.J23117 + I13504 All built into the pSB1C3 plasmid backbone.
In the first week of our iGEM induction, we started out by hydrating DNA from the kit, after which we followed the standard protocols for BioBrick assembly. The first step was transforming our BioBrick DNA into competent DH5α cells (New England Biolabs), and growing them in overnight cultures. After transforming, we performed our first miniprep to obtain the DNA needed for further experiments, and checked the DNA concentration using Qubit. We then used values obtained from this to calculate the volumes of all of components of the digestion step. After the digestion, we ran a part of each sample on a gel to see if the insert and vector were the correct size. The ligation was the last step; after this, we plated our samples and incubated them overnight. We hoped that the next morning, there would be glowing green colonies on our plates. Protocols of these experiments can be found on these pages of the lab book: [transformation, miniprep, Qubit, digestion, ligation]
The next day, we came into the lab, hoping to see colonies so green they would be blinding. After viewing the plates under blue light to visualise the green fluorescent protein (GFP) in our constructs, we understood that the statement ‘biology sometimes doesn’t work’ is entirely accurate. There were no glowing colonies on our plates. However, this moment of truth inspired us to diagnose what was wrong with our devices, and give the Interlab Study another try in the following week.
Our lab induction and Interlab Study were a good chance to find out where the equipment and resources in the lab were, as well as to meet some of the researchers working there – many of them helped at various stages of our project. Those amongst us with little or no lab experience learned basic techniques, such as pouring agar and making LB broth, but also made an effort to understand the biological concepts behind the Interlab Study. One of our physicists, Todd, said that in the first week, he learned the skills necessary for any budding biologist – pipetting, making, and pouring agar. ‘I thought it was very useful. It was great.’ – a direct quote from Todd. Equipped with these skills, we set out to complete the Interlab Study accurately, efficiently, and most importantly – successfully.