Team:Czech Republic/Notebook

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).

Czech Republic M2 gel-05-04.png

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

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

Czech Republic M2 Ste2verif.png

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).

Czech Republic M2 AlphaF1.png

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

Czech Republic M2 AlphaF2.png

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).

Czech Republic M2 pGal.png

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).

Czech Republic pCup-alpha.png

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)

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 (6 microfluidic chips encapsulated on two glass slides)

Week 17 (24-30)

Week 18 (31-06)

Module 5

Signal transmission experiments in microfluidics: MATx + MATa pFUS1-GFP.

September

Week 19 (07-13)

Module 5

Bonding of PDMS to glass (3 microfluidic chips encapsulated on a glass slide)

Signal transmission experiments in microfluidics: MATx + MATa pFUS1-GFP.

Week 20 (14-20)