Team:Tokyo Metropolitan/Notebook/Fiber/2010/09/27
From 2010.igem.org
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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) | ||
- | + | :see [https://2010.igem.org/Team:Tokyo_Metropolitan/Project/Fiber/Protocol#Protocol3:PCR protocol3] | |
- | :''' | + | ===Experiment:Agarose Gels Electrophoresis=== |
+ | |||
+ | :'''Member''' | ||
:Same above | :Same above | ||
Line 29: | Line 31: | ||
: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 39: | Line 41: | ||
: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> | ||
- | + | ===Experiment:Agarose gel Electrophoresis=== | |
- | :''' | + | :'''Member''' |
:Same above | :Same above | ||
Line 47: | Line 49: | ||
:'''Materials and Procedure''' | :'''Materials and Procedure''' | ||
- | :Same | + | :Same <html><a href="https://2010.igem.org/Team:Tokyo_Metropolitan/Project/Fiber/Protocol#.E3.80.80Electrophoresis">protocol4</a></html> |
- | :PCR | + | :PCR Production is the above things. |
- | ===Ligation=== | + | ===Experiment:Ligation=== |
- | :''' | + | :'''Member''' |
:Same above | :Same above | ||
- | :''' | + | :'''Materials and Procedure''' |
- | :Follow | + | :Follow [https://2010.igem.org/Team:Tokyo_Metropolitan/Project/Fiber/Protocol#Protocol8:Ligation protocol8(Ligation)]. |
:*bcsB(E.coli) | :*bcsB(E.coli) | ||
:*bcsC(E.coli) | :*bcsC(E.coli) | ||
- | ===Ligation and Transformation=== | + | ===Experiment: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)=== | + | ===Experimet: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 89: | ||
:(95℃~72℃/30cycle) | :(95℃~72℃/30cycle) | ||
- | ===Agarose Gels | + | ===Experiment:Agarose Gels Electrophoresis=== |
- | :''' | + | :'''Member''' |
:Same above | :Same above | ||
Line 98: | Line 100: | ||
:PCR Product of insert is the above things. | :PCR Product of insert is the above things. | ||
- | ===PCR=== | + | ===Experiment:PCR=== |
+ | :'''Member''' | ||
+ | :watachin and bambi75 | ||
+ | |||
+ | :'''Materials,Procedure''' | ||
:PCR of A.xylinum bcsA,D and E.coli bcsA,B,C ,pSB1C3 | :PCR of A.xylinum bcsA,D and E.coli bcsA,B,C ,pSB1C3 | ||
+ | :Same on August 23th. | ||
+ | :*95℃5min | ||
+ | :*95℃30sec | ||
+ | :*55℃30sec | ||
+ | :*72℃4min | ||
+ | :*72℃5min | ||
+ | :*10℃∞ | ||
+ | :(95℃~72℃/30cycle) | ||
- | ===Electrophoreses of PCR production=== | + | :see [https://2010.igem.org/Team:Tokyo_Metropolitan/Project/Fiber/Protocol#Protocol3:PCR protocol3] |
+ | |||
+ | ===Experiment:Electrophoreses of PCR production=== | ||
+ | |||
+ | :'''Member''' | ||
+ | |||
+ | :Same above | ||
+ | |||
+ | :'''Materials and Procedure''' | ||
+ | |||
+ | :Same protocol on August 16th. | ||
+ | :PCR Product of insert is the above things. | ||
:'''Result''' | :'''Result''' | ||
Line 108: | Line 133: | ||
:[[Image:2010-09-27-01.jpg]] | :[[Image:2010-09-27-01.jpg]] | ||
- | ===Digestion=== | + | ===Experiment:Digestion=== |
:bcsA → XbaI,SpeI | :bcsA → XbaI,SpeI | ||
Line 118: | Line 143: | ||
:pSB1C3 → XbaI, SpeI | :pSB1C3 → XbaI, SpeI | ||
- | ===Electrophoreses of PCR productions=== | + | ===Experiment:Electrophoreses of PCR productions=== |
+ | |||
+ | :'''Member''' | ||
+ | |||
+ | :Same above | ||
+ | |||
+ | :'''Materials and Procedure''' | ||
+ | |||
+ | :Same protocol on August 16th. | ||
+ | :PCR Product of insert is the above things. | ||
:'''Result''' | :'''Result''' | ||
- | [[Image:2010-09-27-02.jpg]] | + | :[[Image:2010-09-27-02.jpg]] |
+ | ===Experiment:DNA Purification=== | ||
+ | :'''member''' | ||
- | + | :naoto and watachin | |
- | + | :'''procedure''' | |
- | + | :see [https://2010.igem.org/Team:Tokyo_Metropolitan/Project/Fiber/Protocol#Protocol6:DNA_Purification_with_silica_gel protocol6] | |
- | ===Ligation=== | + | ===Experiment:Ligation=== |
+ | :'''Member''' | ||
- | + | :Same above | |
- | + | :'''Materials and Procedure''' | |
+ | :Follow [https://2010.igem.org/Team:Tokyo_Metropolitan/Project/Fiber/Protocol#Protocol8:Ligation protocol8(Ligation)]. | ||
- | ===Electrophoreses of PCR productions=== | + | ===Experiment:DNA Purification=== |
+ | :'''member''' | ||
+ | |||
+ | :naoto and watachin | ||
+ | |||
+ | :'''procedure''' | ||
+ | |||
+ | :see [https://2010.igem.org/Team:Tokyo_Metropolitan/Project/Fiber/Protocol#Protocol6:DNA_Purification_with_silica_gel protocol6] | ||
+ | |||
+ | ===Experiment:Ligation=== | ||
+ | :'''Member''' | ||
+ | |||
+ | :Same above | ||
+ | |||
+ | :'''Materials and Procedure''' | ||
+ | :Follow [https://2010.igem.org/Team:Tokyo_Metropolitan/Project/Fiber/Protocol#Protocol8:Ligation protocol8(Ligation)]. | ||
+ | |||
+ | ===Experiment:Transfomation=== | ||
+ | |||
+ | :'''Member''' | ||
+ | :watachin | ||
+ | |||
+ | :'''Materials,Procedure''' | ||
+ | :see [https://2010.igem.org/Team:Tokyo_Metropolitan/Project/Fiber/Protocol#Protocol9:Transformation protocol9] | ||
+ | |||
+ | ===Experiment:PCR=== | ||
+ | :'''Member''' | ||
+ | :watachin | ||
+ | |||
+ | :'''Materials,Procedure''' | ||
+ | :Same on August 23th. | ||
+ | :*95℃5min | ||
+ | :*95℃30sec | ||
+ | :*55℃30sec | ||
+ | :*72℃4min | ||
+ | :*72℃5min | ||
+ | :*10℃∞ | ||
+ | :(95℃~72℃/30cycle) | ||
+ | |||
+ | :see [https://2010.igem.org/Team:Tokyo_Metropolitan/Project/Fiber/Protocol#Protocol3:PCR protocol3] | ||
+ | |||
+ | ===Experiment:Electrophoreses of PCR productions=== | ||
+ | :'''Member''' | ||
+ | |||
+ | :Same above | ||
+ | |||
+ | :'''Materials and Procedure''' | ||
+ | |||
+ | :Same protocol on August 16th. | ||
+ | :PCR Product of insert is the above things. | ||
'''Result''' | '''Result''' | ||
- | [[Image:2010-09-27-03.jpg]] | + | :[[Image:2010-09-27-03.jpg]] |
+ | |||
+ | <br/> |
Latest revision as of 16:55, 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)
- see protocol3
Experiment: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
Experiment:Agarose gel Electrophoresis
- Member
- Same above
- Materials and Procedure
- Same protocol4
- PCR Production is the above things.
Experiment:Ligation
- Member
- Same above
- Materials and Procedure
- Follow protocol8(Ligation).
- bcsB(E.coli)
- bcsC(E.coli)
Experiment:Ligation and Transformation
- Member
- Same above
- Materials and Procedure
- Same protocol on September 21st.
Experimet: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)
Experiment:Agarose Gels Electrophoresis
- Member
- Same above
- Materials and Procedure
- Same protocol on August 16th.
- PCR Product of insert is the above things.
Experiment:PCR
- Member
- watachin and bambi75
- Materials,Procedure
- PCR of A.xylinum bcsA,D and E.coli bcsA,B,C ,pSB1C3
- Same on August 23th.
- 95℃5min
- 95℃30sec
- 55℃30sec
- 72℃4min
- 72℃5min
- 10℃∞
- (95℃~72℃/30cycle)
- see protocol3
Experiment:Electrophoreses of PCR production
- Member
- Same above
- Materials and Procedure
- Same protocol on August 16th.
- PCR Product of insert is the above things.
- Result
Experiment:Digestion
- bcsA → XbaI,SpeI
- bcsB → XbaI
- bcsC → SpeI
- pSB1C3 → XbaI, SpeI
Experiment:Electrophoreses of PCR productions
- Member
- Same above
- Materials and Procedure
- Same protocol on August 16th.
- PCR Product of insert is the above things.
- Result
Experiment:DNA Purification
- member
- naoto and watachin
- procedure
- see protocol6
Experiment:Ligation
- Member
- Same above
- Materials and Procedure
- Follow protocol8(Ligation).
Experiment:DNA Purification
- member
- naoto and watachin
- procedure
- see protocol6
Experiment:Ligation
- Member
- Same above
- Materials and Procedure
- Follow protocol8(Ligation).
Experiment:Transfomation
- Member
- watachin
- Materials,Procedure
- see protocol9
Experiment:PCR
- Member
- watachin
- Materials,Procedure
- Same on August 23th.
- 95℃5min
- 95℃30sec
- 55℃30sec
- 72℃4min
- 72℃5min
- 10℃∞
- (95℃~72℃/30cycle)
- see protocol3
Experiment:Electrophoreses of PCR productions
- Member
- Same above
- Materials and Procedure
- Same protocol on August 16th.
- PCR Product of insert is the above things.
Result