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Team:Tokyo Metropolitan/Notebook/Fiber/2010/09/27
From 2010.igem.org
(Difference between revisions)
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__NOTOC__ | __NOTOC__ | ||
==2010/9/27 Monday (watachin)(bambi75)== | ==2010/9/27 Monday (watachin)(bambi75)== | ||
| - | ===PCR/bcsA and D(A.xylium)/bcsA,B and C(E.coli)=== | + | ===Experiment:PCR/bcsA and D(A.xylium)/bcsA,B and C(E.coli)=== |
| - | :''' | + | :'''Member''' |
:watachin and bambi75 | :watachin and bambi75 | ||
| - | :''' | + | :'''Materials,Procedure''' |
:Same on August 23th. | :Same on August 23th. | ||
:*95℃5min | :*95℃5min | ||
| Line 17: | Line 17: | ||
:（95℃～72℃/30cycle） | :（95℃～72℃/30cycle） | ||
| - | ===Agarose Gels | + | ===Agarose Gels Electrophoresis=== |
| - | :''' | + | :'''Member''' |
:Same above | :Same above | ||
| Line 29: | Line 29: | ||
:PCR Product of insert is the above things. | :PCR Product of insert is the above things. | ||
| - | :=== | + | :===Experiment:Restriction enzyme digestion=== |
| - | :''' | + | :'''Member''' |
:Same above | :Same above | ||
| Line 41: | Line 41: | ||
:===Agarose Gels Electro-phoresis=== | :===Agarose Gels Electro-phoresis=== | ||
| - | :''' | + | :'''Member''' |
:Same above | :Same above | ||
| Line 52: | Line 52: | ||
===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. | ||
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===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 | ||
| Line 87: | Line 87: | ||
:（95℃～72℃/30cycle） | :（95℃～72℃/30cycle） | ||
| - | ===Agarose Gels | + | ===Agarose Gels Electrophoresis=== |
| - | :''' | + | :'''Member''' |
:Same above | :Same above | ||
Revision as of 15:08, 27 October 2010
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 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 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
