In July, we started our lab journey at the Centre for Research in Agricultural Genomics (CRAG). Our first challenge was gene design. We had to optimize our genes to be more compatible with the codon guide of Chlamydomonas reinhardtii, but also to have a low enough content of GC nucleotides so they could be synthesized. To solve this, we had to design a codon optimizer for our genes that took into account both GC content, and the natural codon proportions of Chlamydomonas reinhardtii. All of these improved genes can be found in our Mo-Clo part collection, on the Parts page of our wiki. Eventually, we were able to obtain a compatible sequence, order our genes, and start performing trial transformations. Simultaneously, we were testing non-transformed Chlamydomonas compatibility with nitrate concentrations and wastewater components, to better understand their survival and growth under these conditions; and also to have a control with which to compare our transformed algae’s performance. Last of all, we learned about cytokinin effects, detection, and quantification, which are very intricate time-consuming methods.

Protocols

Lab Notebook