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

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Revision as of 21:23, 26 October 2010

E.coli Fiber Project Notebook

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℃

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