IGEM:Peking/2007/Count-Conjugation-Notebook/2007-7-31: Difference between revisions

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===electrophorsis result===
===electrophorsis result===
*from ''left'' to ''right'':
*from ''left'' to ''right'':
#37℃ @ program 1
#30℃ @ program 1
#37℃ @ program 2
#37℃ @ program 2
#marker


==PCR OriT==
==PCR OriT==

Revision as of 06:37, 1 August 2007

Tandem OriT

  • by Qu Mingzhi
  • get F-OriT primer, primer vf,vf2,vr,pf1,pr1 stored as ~100uM.
  • PCR system contains (totally 50uL):1uL Primer pf1, 1uL Primer pr1, 4uL dNTP, 0.5uL Taq, 1uL OriT template, 37.5uL dH20, 5uL 10X buffer,
  • use different PCR program to test efficiency.
  1. PCR program condition 1: 94℃ 5min, 94℃ 30s, 51℃ 30s, 72℃ 30s, Go to step 2 for 29 times,

72℃ 10min, 4℃ end.

  1. PCR program condition 2: 94℃ 5min, 94℃ 30s, 55℃ 30s, 72℃ 30s, Go to step 2 for 29 times,

72℃ 10min, 4℃ end .

  • Primer final concentration 1uM.

electrophorsis result

  • from left to right:
  1. 37℃ @ program 1
  2. 30℃ @ program 1
  3. 37℃ @ program 2
  4. 37℃ @ program 2
  5. marker

PCR OriT

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