Task:

1. Laboratory safety training.
2. Preparation of LB and YPD solid medium.
3. Autoclaving, pouring of plates, shaking of bacterial solutions.

Task:

1. Extraction of plasmids.
2. PCR amplification of target genes.
3. Electrophoresis to verify PCR results
4. Cut the gel and recover the DNA.

Task:

1. Double cleavage and ligation of the gene fragment to the plasmid backbone
2. Transformation to E. coli DH5α competent cells.

Task:

1. Colony PCR identification of positive transformants.
2. Positive transformants with correct colony PCR results were picked for overnight incubation.

Task:

1. Integrative plasmid digestion to obtain target fragments
2. Agarose electrophoresis for identification and recovery of target fragment gels.

Task: transformation: first round of integration

1. Yeast Cell Expansion Cultures.
2. Preparation of Receptor Yeast Cells.
3. Saccharomyces cerevisiae LiAc transformation.

Task: first round of integration (fail)

1. PCR identification of yeast transformant colonies
2. Expanded culture of yeast transformants with target bands.

Task: Repeat the first round of integration

1. PCR identification of yeast transformant colonies.
2. Expanded culture of yeast transformants with target bands.

Task: second round of integration

1. Preparation of Receptor Yeast Cells.
2. Saccharomyces cerevisiae LiAc transformation.
3. PCR identification of yeast transformant colonies.
4. Expanded culture of yeast transformants with target bands.

Task:

1. Learning the principles of qPCR and how to operate it.
2. Analysis of gene expression in recombinant yeast strains by qPCR.

Task:

1. Shake flask culture of three yeast strains.
2. Ultrasonic crushing of yeast cells.
3. Preparation of β-carotene standard curve.
4. Determination of β-carotene content by microplate reader.

Task:

1. Analyzing experimental data.
2. Writing wiki materials.