Difference between revisions of "Team:Czech Republic/Notebook"

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[[Team:Czech_Republic/Protocols#Transformation_(yeast)|Transformation]]).
 
[[Team:Czech_Republic/Protocols#Transformation_(yeast)|Transformation]]).
  
'''Summary: We successfully transformed EBY100 yeast cells with four different types of yeast display plasmid.
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'''Summary: We successfully transformed EBY100 yeast cells with four different types of yeast display plasmid. We also constructed final BioBrick plasmids.'''
We also constructed final BioBrick plasmids.'''
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=== Module 5 ===
 
=== Module 5 ===

Revision as of 15:37, 11 September 2015

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.

Module 3

Gel verification of pCT302 yeast display plasmid

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.

Module 3

FILLER TEXT

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.

Module 3

FILLER TEXT

Week 4 (25-31)

Module 3

FILLER TEXT

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)

Module 3

FILLER TEXT

Week 15 (10-16)

Module 3

FILLER TEXT

Week 16 (17-23)

Module 3

FILLER TEXT

Module 5

Bonding of PDMS to glass (6 microfluidic chips encapsulated on two glass slides)

Week 17 (24-30)

Module 3

FILLER TEXT

Week 18 (31-06)

Module 3

FILLER TEXT

Module 5

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

September

Week 19 (07-13)

Module 3

PCR amplification of Anti-cMyc, Anti-EpCAM and EpCAM BioBricks from yeast display plasmids (PCR).

Purification of the PCR products, ligation and transformation to DH5alpha bacterial cells (Restriction digest, Ligation, Transformation).

Second attempt of yeast transforamtion with Anti-cMyc, Anti-EpCAM, Anti-HuA and EpCAM inducible yeast display plasmids( Transformation).

Summary: We successfully transformed EBY100 yeast cells with four different types of yeast display plasmid. We also constructed final BioBrick plasmids.

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)