Team:Botchan Lab Tokyo/Notebook
Notebook
Jun 29
• LD medium preparation
July 3
• Nitrogen-free medium A and B preparation
July 5
• Nitrogen-free medium C and D preparation
• glutamate solution and NH4Cl solution preparation
July 20
• ATCC15970 is planted in a LD liquid medium
July 21
• ATCC15970 is planted in a Nitrogen-free medium from LD liquid medium
• Filling of nitrogen
July 24
• Indophenol (72h culture)
"We forgot to take a picture."
July 27
• PCR of ATCC15970 and pUC19
July 28
• Electrophoresis of PCR product
Figure 1 Electrophoresis of PCR product
July 31
• In Fusion Cloning of NifBHDKENXhesA and pUC
August 1
• Transformation
Figure 2 Transformation colony
August 3
• PCR of NBRC 105701 and glu DH,phe DH
August 8
• Insert check PCR of NifBHDKENXhesA
• LB+Km medium preparation
August 9
• Electrophoresis of Insert check PCR product and NBRC 105701,glu DH,phe DH PCR product
Figure 3 Electrophoresis of Insert check PCR product and NBRC 105701,glu DH,phe DH PCR product
August 22
• ATCC15970 is planted in a LB+Amp medium from A
August 23
• Plasmid extraction of transgenic organism
• Electrophoresis of Plasmid extraction product
Figure 4 Electrophoresis of Plasmid extraction product
August 24
• Indophenol
Figure 5 Indophenol
August 25
• Indophenol
Figure 6,7 Indophenol
August 28
• Transgenic organism is planted in a LD liquid medium
August 29
• Plasmid extraction of transgenic organism
• Restriction enzyme treatment of restriction enzyme treatment
August 30
• Ethanol precipitation of restriction enzyme treatment product
• Electrophoresis of restriction enzyme treatment product and no restriction enzyme treatment product
Figure 8 Electrophoresis of restriction enzyme treatment product and no restriction enzyme treatment product
August 31
• Plasmid extraction of transgenic organism
• Electrophoresis of Plasmid extraction product
Figure 9 Electrophoresis of Plasmid extraction product
• Restriction enzyme treatment of restriction enzyme treatment
• Electrophoresis of restriction enzyme treatment product and no restriction enzyme treatment product
Figure 10 Electrophoresis of restriction enzyme treatment product and no restriction enzyme treatment product
September 5
• Insert check PCR of NifBHDKENXhesA
September 6
• Electrophoresis of PCR product
• PCR of Plasmid extraction product
Figure 11 Electrophoresis of PCR product
September 7
• Electrophoresis of PCR product
• PCR of pUC19
Figure 12 Electrophoresis of PCR product
September 8
• Electrophoresis of pUC19 PCR product
Figure 13 Electrophoresis of PCR product
September 18
• ATCC15970 is planted in a LD liquid medium
September 19
• ATCC15970 is planted in a Nitrogen-free medium from LD liquid medium
• Filling of nitrogen
September 20
• Indophenol (24h culture)
September 21
• PCR of pHY300PKL,pUC19 and ATCC15970
September 22
• Electrophoresis of PCR product
Figure 14 Electrophoresis of PCR product
• Indophenol (72h culture)
Figure 15 Indophenol (72h culture)
September 25
• Purification of PCR product
• Electrophoresis of purification product
Figure 16 Electrophoresis of PCR product
September 26
• Electrophoresis of purification product
• PCR of ATCC15970
Figure 17 Electrophoresis of PCR product
September 27
• Electrophoresis of PCR product
Figure 18 Electrophoresis of PCR product
• Purification of PCR product
• Electrophoresis of purification product
Figure 19 Electrophoresis of purification product
September 28
• Transformation
October 2
• Transformation
• ATCC15970 is planted in a LD liquid medium
October 3
• ATCC15970 is planted in a Nitrogen-free medium from LD liquid medium
• Filling of nitrogen
October 4
• Indophenol (24h culture)
Figure 20 Indophenol (24h culture)
October 5
• Indophenol (48h culture)
Figure 21 Indophenol (48h culture)
• In Fusion Cloning of NifBHDKENXhesA and pUC/ NifBHDKENXhesA and pHY
• Transformation
Figure 22,23 Transformation colony
October 6
• Indophenol (72h culture)
Figure 24 Indophenol (72h culture)
• Transformation
Figure 25,26 Transformation colony
October 9
• Preculture of Transformation sample
October 10
• Plasmid extraction
• Electrophoresis of plasmid extraction
Figure 27 Electrophoresis of plasmid extraction
October 12
• Insert check PCR of NifBHDKENXhesA
• Electrophoresis of PCR product
Figure 28 Electrophoresis of PCR product
October 13
• Transformation
October 16
• PCR of NifVSU
• Electrophoresis of PCR product
• Plasmid extraction
Figure 29 Electrophoresis of PCR product
Figure 30 Electrophoresis of Plasmid extraction product
October 17
• Restriction enzyme treatment with Sac1,Xma1
• ATCC15970 and Transformants is planted in a LD liquid medium
October 18
• Ligation of NifVSU and pMW218
• Transformation
Figure 31 Transformation colony
• ATCC15970 and Transformants is planted in a Nitrogen-free medium from LD liquid medium
• Filling of nitrogen
October 19
• Indophenol (24h culture)
Figure 32 Indophenol (24h culture)
• Preculture of Transformation sample
October 20
• Plasmid extraction
• PCR of transformations colony
• Electrophoresis of PCR product
Figure 33 Electrophoresis of PCR product
• Indophenol (48h culture)
Figure 34 Indophenol (48h culture)
October 21
• Electrophoresis of Plasmid extraction product
Figure 35 Electrophoresis of Plasmid extraction product
October 24
• ATCC15970 and Transformants is planted in a LD liquid medium
October 25
• ATCC15970 and Transformants is planted in a Nitrogen-free medium from LD liquid medium
• Filling of nitrogen
October 26
• Indophenol (24h culture)
Figure 36 Indophenol (24h culture)
• ATCC15970 and Transformants is planted in a LD liquid medium
• PCR of transformations colony
October 27
• Electrophoresis of PCR product
Figure 37,38 Electrophoresis of PCR product
• PCR of pheDH and gluDH
• Electrophoresis of PCR product
Figure 39 Electrophoresis of PCR product
• ATCC15970 and Transformants is planted in a Nitrogen-free medium from LD liquid medium
• Filling of nitrogen
• Indophenol (48h culture)
Figure 40 Indophenol (48h culture)
October 30
• Indophenol (72h culture)
Figure 41 Indophenol (72h culture) (without LD medium version)
October 31
• Preculture of Transformation(NifVSU) sample
November 1
• Plasmid extraction
• Electrophoresis of Plasmid extraction product
Figure 42 Electrophoresis of Plasmid extraction product
