Currently we are working hard in our new lab as can be seen in the video. The past couple months we first have been through a very strict lab training given by our supervisors. During the training the supervisors taught us how to work clean, efficiently, and safely with state of the art lab equipments.
After completing the lab training we started our own experiments. The first experiments we started with were cloning experiments in Escherichia coli (E.coli). During the cloning experiments we tried to clone parts of our construct. As many people in the same field may know, this can be quite challenging. Every day we carry out experiments and try to improve them day by day.
Next to the cloning experiments we worked hard to succeed in all the safety issues in order to work with different strain as well. Next to E. coli, we have also started to work with Bacillus Subtilis (B. subtilis), Vibrio Natriegens (V. natriegens) and Pseudomonas putida (P. putida). We are trying to get know how to work with these organisms so we can test our orthogonal plasmid in it.
Moreover, we have received some really useful feedback on our orthogonal replication which we will use to improve our plasmid. Based on the interviews we made with experts in the field of linear plasmid replication, we discovered single and double strength DNA binding proteins in our construct are toxic for our host cells in case we overexpress these proteins. Now we are also busy with experiments in order to characterise the toxicity levels.