Team:Tokyo Metropolitan/Notebook/Fiber/2010/08/12

From 2010.igem.org

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===Experiment:PCR===
===Experiment:PCR===
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'''member'''
+
:'''Member'''
-
NEX , bambi75 and watachin
+
:NEX , bambi75 and watachin
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'''Materials'''
+
:'''Materials'''
-
*sterilized water 106.5μl
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:*sterilized water 106.5μl
-
*Ex taq buffer 15μl
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:*Ex taq buffer 15μl
-
*dNTP 12μl
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:*dNTP 12μl
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*Ex taq 1.5μl
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:*Ex taq 1.5μl
-
*K12bcsA sense(10μmol/l)2.5μl
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:*K12bcsA sense(10μmol/l)2.5μl
-
*K12bcsA antisense(10μmol/l)2.5μl
+
:*K12bcsA antisense(10μmol/l)2.5μl
-
*K12bcsB sense(10μmol/l)2.5μl
+
:*K12bcsB sense(10μmol/l)2.5μl
-
*K12bcsB antisense(10μmol/l)2.5μl
+
:*K12bcsB antisense(10μmol/l)2.5μl
-
*K12bcsC sense(10μmol/l)2.5μl
+
:*K12bcsC sense(10μmol/l)2.5μl
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*K12bcsC antisense(10μmol/l)2.5μl
+
:*K12bcsC antisense(10μmol/l)2.5μl
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*E.coliK12strain
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:*E.coliK12strain
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'''Equipment'''
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:'''Equipment'''
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*thermal cycler
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:*thermal cycler
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*vortex mixer
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:*vortex mixer
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*PCR-tubes
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:*PCR-tubes
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*pipette tip
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:*pipette tip
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'''Procedure'''
+
:'''Procedure'''
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1)mix materials : sterilized water , Ex taq buffer , dNTP and Ex taq.
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:1.mix materials : sterilized water , Ex taq buffer , dNTP and Ex taq and divide 1) into 3 tips.
 +
:2.mix K12bcsAsense , K12bcsAantisense and 2)<br/>
 +
:mix K12bcsBsense , K12bcsBantisense and 2)<br/>
 +
:mix K12bcsCsense , K12bcsCantisense and 2)<br/>
 +
:3.dip E.coliK12strain into 3).
 +
:4.setting tips and elongation in thermal cycler
 +
:*initialization 95℃ 3min
 +
:*denaturation 96℃1min
 +
:*annealing 55℃ 1min
 +
:*elongation 72℃ 5min  (30 cycles from 95℃ to 72℃)
 +
:*reaction stop 10℃
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2)divide 1) into 3 tips.
+
<br/>
-
 
+
-
3)mix K12bcsAsense , K12bcsAantisense and 2).
+
-
 
+
-
　mix K12bcsBsense , K12bcsBantisense and 2).
+
-
 
+
-
　mix K12bcsCsense , K12bcsCantisense and 2).
+
-
 
+
-
4)dip E.coliK12strain into 3).
+
-
 
+
-
5)setting tips and elongation in thermal cycler
+
-
*initialization 95℃ 3min
+
-
*denaturation 96℃1min
+
-
*annealing 55℃ 5min
+
-
*elongation 72℃ 1min  (30 cycles from 95℃ to 72℃)
+
-
*reaction stop 10℃
+

Latest revision as of 21:24, 26 October 2010


E.coli Fiber Project Notebook

August 2010
SUNMONTUEWEDTHUFRISAT
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September 2010
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October 2010
SUNMONTUEWEDTHUFRISAT
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2010/08/12 Tuesday (Bambi75)

Experiment:PCR

Member
NEX , bambi75 and watachin
Materials
  • sterilized water 106.5μl
  • Ex taq buffer 15μl
  • dNTP 12μl
  • Ex taq 1.5μl
  • K12bcsA sense(10μmol/l)2.5μl
  • K12bcsA antisense(10μmol/l)2.5μl
  • K12bcsB sense(10μmol/l)2.5μl
  • K12bcsB antisense(10μmol/l)2.5μl
  • K12bcsC sense(10μmol/l)2.5μl
  • K12bcsC antisense(10μmol/l)2.5μl
  • E.coliK12strain
Equipment
  • thermal cycler
  • vortex mixer
  • PCR-tubes
  • pipette tip
Procedure
1.mix materials : sterilized water , Ex taq buffer , dNTP and Ex taq and divide 1) into 3 tips.
2.mix K12bcsAsense , K12bcsAantisense and 2)
mix K12bcsBsense , K12bcsBantisense and 2)
mix K12bcsCsense , K12bcsCantisense and 2)
3.dip E.coliK12strain into 3).
4.setting tips and elongation in thermal cycler
  • initialization 95℃ 3min
  • denaturation 96℃1min
  • annealing 55℃ 1min
  • elongation 72℃ 5min (30 cycles from 95℃ to 72℃)
  • reaction stop 10℃

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