From 2014.igem.org

Shaping the next generation of scientists

Science is a process of lifelong learning, inspired by curiosity in nature, our surrounding and any unknown processes. The progress in science and technology impacts the daily life of every one, and one of sciences most prominent features, is not only to expand the borders of the human knowledge, but also to teach and educate society. This interplay is important, to enable society to participate in new technologies and progress. Especially new areas of science, such as nanotechnologies and genetic engineering are often met with doubts and fear, which can only be taken away by proper education and providing the necessary information. Synthetic Biology, the intersection of Biology and engineering, is another recent scientific area, which faces skepticism or unawareness. Common points of arguing are related to ethical issues, (Playing God) and the fear of deficient safety regulation and therefore propagation genetic changes outside of the lab. To address these fears, it is important to provide people with biological knowledge, so that they can evaluate the risks for themselves and do not have to depend on second hand knowledge. We thought about providing theoretical and practical skills to young minds, to give them some first hand experience in lab work and the daily life of scientist. Within the Life Science Lab Heidelberg, we offered a 3 day Synthetic Biology lab course for students between 15 and 18 years from high schools in the area of Heidelberg. In these 3 days, students were supposed to repeat the circularization of GFP, an experiment we had previously conduced in the lab (Link to experiment). Additional to the practical background, students we prepared seminars to provide the students with the the basic biological concepts of our experiments, as well as the methods they were using.

Here you can find the protocol we wrote the students which contains all protocols, descriptions and explanation of molecular methods. It is in German language. #

Practical lab work

On the first day, the interns conducted a PCR from a plasmid containing GFP with primers containing BsaI overhangs. The PCR product was visualised on an agarose gel, purified and its concentration determined using a spectroscopical measurement technique. The purified construct was used for golden gate cloning using our toolbox construct for circularization.

The final construct, a positive control containing normal GFP on an expression vector and a negative control, were transformed in BL21 (DE3) and plated on agar containing ampicillin. Additionally liquid cultures were inoculated.

On the second day the liquid cultures were used to inoculate the main expression culture. Moreover a sample was taken for a colony PCR to verify the presence of our circularization construct and the linear GFP construct.

Upon the ID of 0.8 the school interns induced the cultures with 1 mM IPTG for one hour. To visualise the proteins- linear and circular GFP version- an SDS- PAGE and Western Blot was conducted.

If our construct leads to the circularization of GFP a shift should be visible on the Western Blot between linear and circular protein. The circular construct runs faster on a gel, since its coiled nature experiences less resistance from the gel matrix.

Unfortunately, the pupils experienced some difficulties in the experiments. To conduct an experiment properly it needs a lot of attention and at least some experience. Even though the results did not look like expected or desired, the pupils learned many new methods and quite some theoretical background. Altogether not only the school kids had much fun with their first-hand experience in the lab, but also we as supervisors and tutors.