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

From 2010.igem.org

(Difference between revisions)
Line 4: Line 4:
===PCR/bcsA and D(A.xylium)/bcsA,B and C(E.coli)===
===PCR/bcsA and D(A.xylium)/bcsA,B and C(E.coli)===
-
'''member'''
+
:'''member'''
-
watachin and bambi75
+
:watachin and bambi75
-
'''materials,Procedure'''
+
:'''materials,Procedure'''
-
Same on August 23th.
+
:Same on August 23th.
-
*95℃5min
+
:*95℃5min
-
*95℃30sec
+
:*95℃30sec
-
*55℃30sec
+
:*55℃30sec
-
*72℃4min
+
:*72℃4min
-
*72℃5min
+
:*72℃5min
-
*10℃∞
+
:*10℃∞
-
（95℃～72℃/30cycle）
+
:（95℃～72℃/30cycle）
===Agarose Gels Electro-phoresis===
===Agarose Gels Electro-phoresis===
-
'''member'''
+
:'''member'''
-
Same above
+
:Same above
-
'''Materials and Procedure'''
+
:'''Materials and Procedure'''
-
Same protocol <html><a href="https://2010.igem.org/Team:Tokyo_Metropolitan/Project/Fiber/Protocol#.E3.80.80agarose gel electrophoresis">the protocol4</a></html>
+
:Same protocol <html><a href="https://2010.igem.org/Team:Tokyo_Metropolitan/Project/Fiber/Protocol#.E3.80.80agarose gel electrophoresis">the protocol4</a></html>
-
PCR Product of insert is the above things.
+
:PCR Product of insert is the above things.
-
===restriction enzyme digestion===
+
:===restriction enzyme digestion===
-
'''member'''
+
:'''member'''
-
Same above  
+
:Same above  
-
'''Materials and Procedure'''
+
:'''Materials and Procedure'''
-
Same protocol <html><a href="https://2010.igem.org/Team:Tokyo_Metropolitan/Project/Fiber/Protocol#.E3.80.80restriction enzyme digestion">the protocol6</a></html>
+
:Same protocol <html><a href="https://2010.igem.org/Team:Tokyo_Metropolitan/Project/Fiber/Protocol#.E3.80.80restriction enzyme digestion">the protocol6</a></html>
 +
:===Agarose Gels Electro-phoresis===
-
===Agarose Gels Electro-phoresis===
+
:'''member'''
-
'''member'''
+
:Same above
-
Same above
+
:'''Materials and Procedure'''
-
'''Materials and Procedure'''
+
:Same protocol on August 16th.
-
 
+
:PCR Product of insert is the above things.
-
Same protocol on August 16th.
+
-
PCR Product of insert is the above things.
+
===Ligation===
===Ligation===
-
'''member'''
+
:'''member'''
-
Same above
+
:Same above
-
'''materials and procedure'''
+
:'''materials and procedure'''
-
Follow the protocol(Ligation).
+
:Follow the protocol(Ligation).
-
*bcsB(E.coli)
+
:*bcsB(E.coli)
-
*bcsC(E.coli)
+
:*bcsC(E.coli)
===Ligation and Transformation===
===Ligation and Transformation===
-
'''member'''
+
:'''member'''
-
Same above
+
:Same above
'''materials and procedure'''
'''materials and procedure'''
-
Same protocol on September 21st.
+
:Same protocol on September 21st.
===PCR/bcsQ and bcsEFG(E.coli)===
===PCR/bcsQ and bcsEFG(E.coli)===
-
'''member'''
+
:'''member'''
-
watachin and bambi75
+
:watachin and bambi75
-
'''materials,Procedure'''
+
:'''materials,Procedure'''
-
Same on August 23th.
+
:Same on August 23th.
-
*95℃5min
+
:*95℃5min
-
*95℃30sec
+
:*95℃30sec
-
*55℃30sec
+
:*55℃30sec
-
*72℃4min
+
:*72℃4min
-
*72℃5min
+
:*72℃5min
-
*10℃∞
+
:*10℃∞
-
（95℃～72℃/30cycle）
+
:（95℃～72℃/30cycle）
===Agarose Gels Electro-phoresis===
===Agarose Gels Electro-phoresis===
-
'''member'''
+
:'''member'''
-
Same above
+
:Same above
-
'''Materials and Procedure'''
+
:'''Materials and Procedure'''
-
Same protocol on August 16th.
+
:Same protocol on August 16th.
-
PCR Product of insert is the above things.
+
:PCR Product of insert is the above things.
===PCR===
===PCR===
-
PCR of A.xylinum bcsA,D and E.coli bcsA,B,C ,pSB1C3
+
 
 +
:PCR of A.xylinum bcsA,D and E.coli bcsA,B,C ,pSB1C3
 +
 
===Electrophoreses of PCR production===
===Electrophoreses of PCR production===
-
'''Result'''
 
-
[[Image:2010-09-27-01.jpg]]
+
:'''Result'''
 +
 
 +
:[[Image:2010-09-27-01.jpg]]
===Digestion===
===Digestion===
-
bcsA → Xba,Spe
 
-
bcsB → Xba
+
:bcsA → XbaI,SpeI
 +
 
 +
:bcsB → XbaI
-
bcsC → Spe
+
:bcsC → SpeI
-
pSB1C3 → Xba, Spe
+
:pSB1C3 → XbaI, SpeI
===Electrophoreses of PCR productions===
===Electrophoreses of PCR productions===
-
'''Result'''
+
:'''Result'''
[[Image:2010-09-27-02.jpg]]
[[Image:2010-09-27-02.jpg]]

Revision as of 15:03, 27 October 2010


E.coli Fiber Project Notebook

August 2010
SUNMONTUEWEDTHUFRISAT
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15 16 17 18 19 20 21
22 23 24 25 26 27 28
29 30 31
September 2010
SUNMONTUEWEDTHUFRISAT
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19 20 21 22 23 24 25
26 27 28 29 30
October 2010
SUNMONTUEWEDTHUFRISAT
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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

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 → 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
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