Team:TMU-Tokyo/Notebook experiment/1st

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Revision as of 17:54, 27 September 2013



Experiment > Parts Synthesis > 1st week



September 2nd
PCR following fragment. (1st PCR)
FragmentPtetR1PtetR2FLPSmaraC-P
PrimerCmNR-stu
FLP-IRR/IRL
CmCR-stu
FLP-IRR/IRL
FLP-L
FLP-R
FLP-SaApL
CmC
acaC-2
BAD-C3
TempleteIRR/IRL-GFP
(BBa_K880002)
IRR/IRL-GFP
(BBa_K880002)
pFTGTm(C2-Ap)SmpBAD/Myc-HisA

FragmenthbiFTclacIFRT-CmFRT-Ap
PrimerpBAD-HbiF
RTcN-hbiF
TcN
TcC3
141-70
FRT-lacIC
R6Kori-Xho-3R
R(FRT)2
lacI-R(FRT)2
mhpA-SaApR
TempletehbiF
(BBa_K880000)
Escherichia coli
MG1655 recA⁻
(cell suspention)
Escherichia coli
MG1655 genome
pSG76CpSG76A2
Result

1: PterR1, 2: PtetR2, 3: FLP, 4: Sm, 5: araC-P,
6: hbiF, 7: Tc, 8: LacI, 9: FRT-Cm, 10: FRT-Ap


September 3rd
Vector Remodeling with PCR. Insertion Recognition sites (Blunt).
FragmentKm-Blunt
PrimerKm-Blunt-N
Km-Blunt-C
TempletepACYC177
Result



DNA Existract following fragment from gel.
PtetR1, FLP, Sm, araC-P, hbiF, Tc, lacI, FRT-Cm, FRT-Ap, Km-Blunt


September 4th
PCR following fragment. (2nd PCR)
FragmentFragment 1Fragment 2Fragment 3
PrimerlacA-IRR/IRL
cynR-SaApL
araCHC
araA-TcC
R(SaApR)-lacIN
mhpA-CmC6
TempletePtetR
FLP
Sm
araC-P
hbiF
Tc
lacI
FRT-Cm
Result

1: Fragment 1, 2: Fragment 2, 3: Fragment 3


September 5th
PCR following fragment. (2nd PCR)
FragmentFragment 1-AFragment 1-BFragment 2-CFragment 2-D
PrimerCmNR-Stu
FLP-R
FLP-L
CmC
araC2
RTcN-hbiF
pBAD-hbiF
TcC3
TempletePtetR
FLP
FLP
Sm
araC-P
hbiF
hbiF
Tc
Result

1: Fragment 1-A, 2: Fragment 1-B,
3: Fragment 2-C, 4: Fragment 2-D


Check following gel extract product.
PtetR1, FLP, Sm, araC-P, hbiF, Tc, lacI, FRT-Cm, FRT-Ap, Km-Blunt
Result

1: PtetR1, 2: FLP, 3: Sm, 4: araC-P, 5: hbiF,
6: Tc, 7: lacI, 8: FRT-Cm, 9: FRT-Ap, 10: Km-Blunt


September 6th
PCR following fragment. (2nd PCR)
FragmentFragment 1-AFragment 1-BFragment 2-AFragment 2-B
PrimerCmNR-Stu
FLP-R
FLP-L
CmC
FLP-L
CmC
FLP-L
cynR-SaApL
TempletePtetR
FLP
FLP
Sm
Sm
FLP
Sm
FLP
Result

1: Fragment 1-A, 2: Fragment 2-B
(1-B and 2-A was failed in PCR)


PCR following fragment. (2nd PCR)
FragmentFragment IFFragment FS
PrimerlacA-IRR/IRL
FLP-R
FLP-L
cynR-SaApR
TempletePtetR
FLP
FLP
Sm


September 7th
PCR Result (Fragment IF, FS)

1: Fragment IF, 2: Fragment FS


PCR following fragment. (3rd PCR)
FragmentFragment 1Fragment 2
PrimerlacA-IRR/IRL
cynR-SaApR
araCHC
araA-TcC
TempleteFragment IF
Fragment FS
Fragment 2-B
Fragment 2-C
Result

1: Fragment 1, 2: Fragment 2


>>Go to next week