Notebook

May

Week 1 (4-10)

Module 2

PCR of the ste2 gene from the genome of Saccharomyces cerevisiae 7284 strain (Genome extraction, PCR, Gel electrophoresis).

Cloning of the amplified ste2 into pRSII416 vector (Restriction digest, Ligation, Transformation).

Verification of the pRSII416_ste2 construct (Miniprep, Restriction digest, Gel electrophoresis).

Summary: We successfully obtained ste2 gene from the yeast genome and cloned it into pRSII416 vector.

Week 2 (11-17)

Module 2

PCR of the incomplete MF(ALPHA)1 locus (see the Design page for details) from the genome of Saccharomyces cerevisiae 7283 strain (Genome extraction, PCR, Gel electrophoresis).

Band-stab PCR of the amplified incomplete MF(ALPHA)1 locus in order to add the actual pheromone and a stop codon.

Cloning of the obtained pheromone into pRSII416 vector (Restriction digest, Ligation, Transformation).

Verification of the pRSII416_sc_alpha construct (Miniprep, Restriction digest, Gel electrophoresis).

Summary: We successfully obtained mating pheromone from Saccharomyces cerevisiae with its secretion tag and cloned it into pRSII416 vector. We also prepared pCUP1 promoter for the cloning upstream of this pheromone.

Week 3 (18-24)

Module 2

PCR amplification of pGAL1 promoter from the genome of Saccharomyces cerevisiae 7284 strain (PCR, Gel electrophoresis).

Cloning of the amplified pCUP1 from Week 2 into pRSII416_sc_alpha plasmid (Restriction digest, Ligation, Transformation).

Verification of the pRSII416_pCUP1_sc_alpha construct (Miniprep, Restriction digest, Gel electrophoresis).

Summary: We have successfully obtained plasmid with a mating pheromone under the control of pCUP1 promoter and prepared pGAL1 promoter for the control of ste2 gene.

Week 4 (25-31)

June

Week 5 (1-07)

Week 6 (08-14)

Week 7 (15-21)

Week 8 (22-28)

Week 9 (29-05)

Module 5

PDMS molding (40μm high silicon masters, 40g PDMS, curing 80°C/2h)

July

Week 10 (06-12)

Week 11 (13-19)

Week 12 (20-26)

Week 13 (27-02)

August

Week 14 (03-09)

Week 15 (10-16)

Week 16 (17-23)

Module 5

Bonding of PDMS to glass (2.5 minutes air plasma treatment, post-bake 80°C/1h)

Week 17 (24-30)

Week 18 (31-06)

September

Week 19 (07-13)

Module 5

Bonding of PDMS to glass (2.5 minutes air plasma treatment, post-bake 80°C/1h)

Week 20 (14-20)