Team:TMU-Tokyo/Notebook/Experiment/4th week (9. 3 - 9. 9)

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■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

 

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