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

E.coli Fiber Project Notebook

August 2010 SUN MON TUE WED THU FRI SAT 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31

September 2010 SUN MON TUE WED THU FRI SAT 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30

October 2010 SUN MON TUE WED THU FRI SAT 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31

Contents 1 2010/9/27 Monday (watachin)(bambi75) 1.1 PCR/bcsA and D(A.xylium)/bcsA,B and C(E.coli) 1.2 Agarose Gels Electro-phoresis 2 restriction enzyme digestion 2.1 member 2.2 Materials and Procedure 3 Agarose Gels Electro-phoresis 3.1 member 3.2 Materials and Procedure 4 Ligation 4.1 member 4.2 materials and procedure 5 Ligation and Transformation 5.1 member 5.2 materials and procedure 6 PCR/bcsQ and bcsEFG(E.coli) 6.1 member 6.2 materials,Procedure 7 Agarose Gels Electro-phoresis 7.1 member 7.2 Materials and Procedure 7.3 PCR 7.4 Electrophoreses of PCR production 7.5 Digestion 7.6 Electrophoreses of PCR productions 7.7 DNA Purification 7.8 Ligation 7.9 DNA Purification 7.10 Ligation 7.11 Transfomation 7.12 PCR 7.13 Electrophoreses of PCR productions

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

Digestion

bcsA → Xba,Spe

bcsB → Xba

bcsC → Spe

pSB1C3 → Xba, Spe

Electrophoreses of PCR productions

Result

DNA Purification

Ligation

DNA Purification

Ligation

Transfomation

PCR

Electrophoreses of PCR productions

Result