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

From 2010.igem.org

(Difference between revisions)
Line 5: Line 5:
'''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
Line 24: Line 26:
*vortex mixer
*vortex mixer
*PCR-tubes
*PCR-tubes
-
*pipet tip
+
*pipette tip
'''Procedure'''
'''Procedure'''

Revision as of 16:25, 24 October 2010


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.

2)divide 1) into 3 tips.

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℃