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

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E.coli Fiber Project Notebook

August 2010
SUNMONTUEWEDTHUFRISAT
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September 2010
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October 2010
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31

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

2010-09-27-01.jpg

Digestion

bcsA → Xba,Spe

bcsB → Xba

bcsC → Spe

pSB1C3 → Xba, Spe

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