Protocols

This section compiles and describes in detail the experimental protocols used throughout the development of the project. These procedures were selected, adapted, or designed based on scientific literature, technical guidelines from specialized laboratories, and the team's previous experience.

Each protocol has been standardized and tested to ensure the reproducibility of the results obtained during the experimental work. The objective of this section is to facilitate the understanding and replication of the experimental work, both for other iGEM teams and for anyone interested in applying or improving these methodologies.

Experimental Protocols (PDF)

• 1. Activation GS115
• 2. Yeast Genomic DNA Isolation using Wizard® Genomic DNA Purification Kit (Promega)
• 3. Wizard® Genomic DNA Purification Kit Promega — Isolation of genomic DNA from bacteria
• 4. DNA extraction with Bioline KIT
• 5. Extraction of genomic DNA from yeast
• 6. Preparation of LB Medium with 5 g/L Gellan Gum Powder
• 7. Laboratory sterilization using instant pot pressure cooker
• 8. Propagating E. coli DH5α cells transformed with pPICZAα
• 9. Miniprep Plasmid DNA Extraction from E. coli
• 10. Activation and propagation of the OpenPichia pep4 yps1 strain
• 11. Glycerol stocks of the OpenPichia pep4 yps1 and E. coli DH5α strains transformed with pPICZAα
• 12. Pre-inoculation
• 13. Inoculation
• 14. Determination of the growth rate