Team:Tokyo Metropolitan/Notebook/Fiber/2010/08/26

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{{:Team:Tokyo_Metropolitan/Header}}
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{{:Team:Tokyo_Metropolitan/Notebook/Fiber}}
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==2010/08/26(Bambi75)==
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==2010/08/26 Thursday(Bambi75)==
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==Make Plates==
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===Experiment:PCR===
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===member===
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:'''Member'''
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NEX , Bambi75 and watachin
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===Materials===
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:NEX , Bambi75 and watachin
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*RO water 200ml
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*LB Broth 4g
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*Cam(50μg/L) 10ml
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===Procedure===
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:'''Materials'''
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① mix materials.
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② Divide ①equally and make 10 plates.
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:*2×PCR buffer  25×4μl
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:*2mM dNTP  10×4μl
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:*10mM primer(sense)bcsA,B,C and D 2.5μl each
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:*10mM primer(antisense)bcsA,B,C and D 2.5μl each
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:*template DNA a little
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:*Q water 9×4μl
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:*KOD FX 0.5×4μl
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==Separation of A.xylinum==
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:'''Procedure'''
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===member===
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:①mix all materials for 4 tubes.
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easily and naoto
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===Materials===
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:②elongation
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*the plate(made on August 25th).
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:*bcsA,bcsB and bcsC
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:**94℃ 2min
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:**98℃ 10sec☆
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:**55℃ 30sec
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:**68℃ 4min★
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:**68℃ 7min
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:**10℃　∞
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:*bcsD
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:**94℃ 2min
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:**98℃ 10sec☆
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:**55℃ 30sec
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:**68℃ 1min★
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:**68℃ 7min
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:**10℃　∞
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:※30cycle ☆ to ★.
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===Procedure===
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<br/>
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①extract material 2ml.
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②centrifuge ① 10000rpm/5min.
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==PCR==
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===member===
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same above
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-
 
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===Materials===
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*2×PCR buffer  25×4μl
+
-
*2mM dNTP  10×4μl
+
-
*10mM primer(sense)bcsA,B,C and D 2.5μl each
+
-
*10mM primer(antisense)bcsA,B,C and D 2.5μl each
+
-
*template DNA a little
+
-
*Q water 9×4μl
+
-
*KOD FX 0.5×4μl
+
-
 
+
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===Procedure===
+
-
①mix all materials for 4 tubes.
+
-
 
+
-
②elongation
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-
*bcsA,bcsB and bcsC
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-
**94℃ 2min
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-
**98℃ 10sec☆
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**55℃ 30sec
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**68℃ 4min★
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**68℃ 7min
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**10℃　∞
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*bcsD
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**94℃ 2min
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**98℃ 10sec☆
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**55℃ 30sec
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-
**68℃ 1min★
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**68℃ 7min
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**10℃　∞
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※30cycle ☆ to ★.
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-
 
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==PCR==
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===member===
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NEX , Bambi75 and watachin
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-
 
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===Materials===
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*sterilized water 71μl
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*Ex taq buffer 10μl
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*dNTP mix 8μl
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*Ex taq 1μl
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*primer(bcsC sense/antisense) 5μl each
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===Procedure===
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①mix materials
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②divide ① into 2 tubes.
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③elongation
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**95℃ 3min
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**96℃ 1min☆
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**55℃ 7min
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**72℃ 1min★
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**10℃ ∞
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※30 cycle ☆to★.
+

Latest revision as of 13:56, 27 October 2010


E.coli Fiber Project Notebook

August 2010
SUNMONTUEWEDTHUFRISAT
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September 2010
SUNMONTUEWEDTHUFRISAT
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October 2010
SUNMONTUEWEDTHUFRISAT
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31

2010/08/26 Thursday(Bambi75)

Experiment:PCR

Member
NEX , Bambi75 and watachin
Materials
  • 2×PCR buffer 25×4μl
  • 2mM dNTP 10×4μl
  • 10mM primer(sense)bcsA,B,C and D 2.5μl each
  • 10mM primer(antisense)bcsA,B,C and D 2.5μl each
  • template DNA a little
  • Q water 9×4μl
  • KOD FX 0.5×4μl
Procedure
①mix all materials for 4 tubes.
②elongation
  • bcsA,bcsB and bcsC
    • 94℃ 2min
    • 98℃ 10sec☆
    • 55℃ 30sec
    • 68℃ 4min★
    • 68℃ 7min
    • 10℃　∞
  • bcsD
    • 94℃ 2min
    • 98℃ 10sec☆
    • 55℃ 30sec
    • 68℃ 1min★
    • 68℃ 7min
    • 10℃　∞
※30cycle ☆ to ★.

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