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

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E.coli Fiber Project Notebook

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