| Abstract [eng] |
The main objective of this master thesis was to identify the optimal transformation by integrating plasmids conditions of S. cerevisiae SP1 strain cells using a pulsed electric field. In order to reach this objective certain experiments were conducted, including, but not limited to cultivation of microorganisms, genotyping, transformation, and visualisation. After transforming S. cerevisiae SP1 strain cells the optimal conditions were found to be 10 kV/cm, 5 × 0,5 ms. The achieved transformation efficiency was 1 × 10^3 transformants/μg DNR. The plasmid was succesfully linearised using a restriction endonuclease StuI (also known as Eco147I). It has been determined that it most appropriate to use early log phase cells and a final cell density of OD600 – 0,91. |
