Team ice-breaking
Project Discussion
Team:SHSID/Notebook
NOTEBOOK
6/26
6/27
Laboratory Safety Training
Media preparation
Transfer and culture E. coli and the yeast
PCR to amplify the target genes, AsADC、speB and pYES2
Transfer and culture E. coli and the yeast
PCR to amplify the target genes, AsADC、speB and pYES2
6/28
Plasmid Extraction: pYES2-X-1-XI-4
PCR verification
PCR verification
6/29
One-day off
6/30
Prepare the competent cells of the yeast
PCR to amplify the vector
PCR to amplify the vector
7/1
Repeat PCR
7/2
Overlap PCR
PCR verification
PCR verification
2kM 1 2 3 4 5 6 7 8
All lanes are correct.
7/3
Team Photo Shooting
DNA gel extraction
Overlap PCR
DNA gel extraction
Overlap PCR
2kM 1 2 3 4
All lanes are correct.
7/4
Overlap PCR
PCR verification
PCR verification
15kM 1 2 3 4 5 6 2kM
Gel electrophoresis diagram. Lane 1, 3, and 5 represent DNA segment 45678, and
lane 2, 4, and 6 represent segment 678.
7/5
KOD FX PCR to amplify the terminator CYC1
Transformation into E. coli
Transformation into E. coli
7/6
Check the transformants
Repeat the transformation
Repeat the transformation
7/7
Repeat: KOD FX PCR to amplify the terminator CYC1
Overlap PCR
Transformation
Overlap PCR
Transformation
7/8
Colony PCR
September
Complete the plasmid construction
Send for sequencing
Transformation into the yeast
10/1-10.7
Fermentation test
Send out the samples for measuring the putrescine yield by LC-MS/MS
Send out the samples for measuring the putrescine yield by LC-MS/MS