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Team:Tokyo Metropolitan/Notebook/Fiber/2010/09/27
From 2010.igem.org
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===restriction enzyme digestion=== | ===restriction enzyme digestion=== | ||
| - | + | '''member''' | |
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Same above | Same above | ||
| - | + | ||
| + | '''Materials and Procedure''' | ||
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Same protocol <html><a href="https://2010.igem.org/Team:Tokyo_Metropolitan/Project/Fiber/Protocol#.E3.80.80restriction enzyme digestion">the protocol6</a></html> | Same protocol <html><a href="https://2010.igem.org/Team:Tokyo_Metropolitan/Project/Fiber/Protocol#.E3.80.80restriction enzyme digestion">the protocol6</a></html> | ||
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===Agarose Gels Electro-phoresis=== | ===Agarose Gels Electro-phoresis=== | ||
| - | + | '''member''' | |
| + | |||
Same above | Same above | ||
| - | + | ||
| + | '''Materials and Procedure''' | ||
| + | |||
Same protocol on August 16th. | Same protocol on August 16th. | ||
PCR Product of insert is the above things. | PCR Product of insert is the above things. | ||
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===Ligation=== | ===Ligation=== | ||
| - | + | '''member''' | |
| + | |||
Same above | Same above | ||
| - | + | ||
| + | '''materials and procedure''' | ||
Follow the protocol(Ligation). | Follow the protocol(Ligation). | ||
*bcsB(E.coli) | *bcsB(E.coli) | ||
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===Ligation and Transformation=== | ===Ligation and Transformation=== | ||
| - | + | '''member''' | |
| + | |||
Same above | Same above | ||
| - | + | '''materials and procedure''' | |
| + | |||
Same protocol on September 21st. | Same protocol on September 21st. | ||
===PCR/bcsQ and bcsEFG(E.coli)=== | ===PCR/bcsQ and bcsEFG(E.coli)=== | ||
| - | + | '''member''' | |
| + | |||
watachin and bambi75 | watachin and bambi75 | ||
| - | + | '''materials,Procedure''' | |
Same on August 23th. | Same on August 23th. | ||
*95℃5min | *95℃5min | ||
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===Agarose Gels Electro-phoresis=== | ===Agarose Gels Electro-phoresis=== | ||
| - | + | '''member''' | |
| + | |||
Same above | Same above | ||
| - | |||
| - | |||
| - | |||
| + | '''Materials and Procedure''' | ||
| + | Same protocol on August 16th. | ||
| + | PCR Product of insert is the above things. | ||
===PCR=== | ===PCR=== | ||
Revision as of 17:47, 24 October 2010
E.coli Fiber Project Notebook
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2010/9/27 Monday (watachin)(bambi75)
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 Electro-phoresis
member
Same above
Materials and Procedure
Same protocol the protocol4
PCR Product of insert is the above things.
restriction enzyme digestion
member
Same above
Materials and Procedure
Same protocol the protocol6
Agarose Gels Electro-phoresis
member
Same above
Materials and Procedure
Same protocol on August 16th. PCR Product of insert 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 Electro-phoresis
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 → Xba,Spe
bcsB → Xba
bcsC → Spe
pSB1C3 → Xba, Spe
Electrophoreses of PCR productions
Result
DNA Purification
Ligation
DNA Purification
Ligation
Transfomation
PCR
Electrophoreses of PCR productions
Result
