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Team:TMU-Tokyo/Notebook/Experiment/4th week (9. 3 - 9. 9)
From 2012.igem.org
■Experiment
1st week (8.13 - 8.19)
2nd week (8.20 - 8.26)
3rd week (8.27 - 9. 2)
4th week (9. 3 - 9. 9)
5th week (9.10 - 9.16)
6th week (9.17 - 9.23)
7th week (9.24 - 9.30)
Experiment
■4th week (9. 3 - 9. 9)
■3rd September
STEP 1: Construction of Device3
PCR of insert (fdh4AB)
Electrophoresis
Failure: because of too dilute bands
PCR of insert (fdh4AB)
Electrophoresis
Failure: because of annealing temperature
PCR of insert (fdh4AB)
Electrophoresis
Success: We observed appropriate bands!
■ 4th Sep
STEP 2: Construction of Device1
Colony PCR of transformed E. coli (pfrm-frmR-GFP)
Electrophoresis
Failure: because of no bands
■ 5th Sep
Ordered P.putida arrive
STEP 2: Construction of Device1
Electrophoresis
Failure: because of no bands
Colony PCR of transformed E. coli (pfrm-frmR-GFP)
Electrophoresis
Success: 2 of 15 samples are appropriate bands!
Electrophoresis
Failure: because of transformation miss
Electrophoresis
Failure: because of annealing temperature
Electrophoresis
Failure: because of annealing temperature
Electrophoresis
Success: 1 of 15 samples is appropriate band!
■ 6th Sep
STEP 2: Construction of Device1
Colony PCR of transformed E. coli (pfrm-frmR-GFP)
Electrophoresis
Failure: because of transformation miss
Electrophoresis
Failure: because of transformation miss
STEP 1: Construction of Device3
Colony PCR of transformed E. coli (p-fdh4AB)
Electrophoresis
Failure: because of no bands
■ 7th Sep
STEP 1: Construction of Device3
PCR of insert (fdh4AB)
Electrophoresis
Success: We observed appropriate bands but a lot of smear.
Electrophoresis
Success: We observed appropriate bands!
Colony PCR of transformed E. coli (p-fdh4AB)
Electrophoresis
Failure: because of PCR condition or ligation miss
STEP 3: Construction of Device2
Preparation of culture plates for P.putida (FDH)
Cultivation of P.putida (FDH) on plate
■ 9th Sep
STEP 3: Construction of Device2
PCR of insert (FDH)
Electrophoresis
Failure: because of no bands
