From 2014.igem.org

Shaping the next generation of scientists

The progress in natural sciences and technology has an impact on the life of each and every one within a society. Therefore it is enormously important to raise discussions in close dialogue with the general public about ethical issues, risks and benefits in Synthetic Biology.

To participate in considerations and evaluations about Synthetic Biology, a basic comprehension of biological processes is essential. To spread knowledge about the practical handling of Synthetic Biology within an actual lab, we offered a three days practical course to pupils at the age of 15 to 18 within the Liege Science Lab in Heidelberg. The Heidelberg Life-Science Lab is part of the German Cancer Research Center (DKFZ). It is dedicated to support extraordinarily gifted and talented high-school students in developing their scientific as well as personal skills and in gaining practical research experience. The school students actually took over the experiments we were performing in the lab and tried to circularize GFP. The fact that our young scientist produced data, which were actually very crucial to us, motivated them to try very hard for the best results. Alternating in theoretical lectures and practical courses, the pupils learned the background of our project and the actual experiments first-hand:

¶protocol

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.