Team:Tokyo Metropolitan/Notebook/Fiber/2010/09/27

From 2010.igem.org

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===Experiment:Restriction enzyme digestion===
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Revision as of 15:17, 27 October 2010


E.coli Fiber Project Notebook

August 2010
SUNMONTUEWEDTHUFRISAT
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September 2010
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October 2010
SUNMONTUEWEDTHUFRISAT
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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
2010-09-27-01.jpg

Digestion

bcsA → XbaI,SpeI
bcsB → XbaI
bcsC → SpeI
pSB1C3 → XbaI, SpeI

Electrophoreses of PCR productions

Result

2010-09-27-02.jpg


DNA Purification

Ligation

DNA Purification

Ligation

Transfomation

PCR

Electrophoreses of PCR productions

Result

2010-09-27-03.jpg