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

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Revision as of 15:17, 27 October 2010

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）

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

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

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 :PCR Product of insert is the above things.
-:===Experiment:Restriction enzyme digestion===
+===Experiment:Restriction enzyme digestion===
 :'''Member'''