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Team:Tokyo Metropolitan/Notebook/Fiber/2010/09/27
From 2010.igem.org
E.coli Fiber Project Notebook
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2010/9/27 Monday (watachin)(bambi75)
Experiment:PCR/bcsA and D(A.xylium)/bcsA,B and C(E.coli)
- Member
- watachin and bambi75
- Materials,Procedure
- Same on August 23th.
- 95℃5min
- 95℃30sec
- 55℃30sec
- 72℃4min
- 72℃5min
- 10℃∞
- （95℃～72℃/30cycle）
Agarose Gels Electrophoresis
- Member
- Same above
- Materials and Procedure
- Same protocol the protocol4
- PCR Product of insert is the above things.
Experiment:Restriction enzyme digestion
- Member
- Same above
- Materials and Procedure
- Same protocol the protocol6
Agarose gel Electrophoresis
- Member
- Same above
- Materials and Procedure
- Same protocol4
- PCR Production is the above things.
Ligation
- Member
- Same above
- Materials and Procedure
- Follow the protocol(Ligation).
- bcsB(E.coli)
- bcsC(E.coli)
Ligation and Transformation
- Member
- Same above
Materials and Procedure
- Same protocol on September 21st.
PCR/bcsQ and bcsEFG(E.coli)
- Member
- watachin and bambi75
- Materials,Procedure
- Same on August 23th.
- 95℃5min
- 95℃30sec
- 55℃30sec
- 72℃4min
- 72℃5min
- 10℃∞
- （95℃～72℃/30cycle）
Agarose Gels Electrophoresis
- Member
- Same above
- Materials and Procedure
- Same protocol on August 16th.
- PCR Product of insert is the above things.
PCR
- PCR of A.xylinum bcsA,D and E.coli bcsA,B,C ,pSB1C3
Electrophoreses of PCR production
- Result
Digestion
- bcsA → XbaI,SpeI
- bcsB → XbaI
- bcsC → SpeI
- pSB1C3 → XbaI, SpeI
Electrophoreses of PCR productions
- Result
DNA Purification
Ligation
DNA Purification
Ligation
Transfomation
PCR
Electrophoreses of PCR productions
Result
