Sentelab:Notebook/gene synthesis/2010/09/17: Difference between revisions

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#4C unlimited time
#4C unlimited time


*results
[[Image:1.pcr products.jpg]]


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Revision as of 05:24, 17 September 2010

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gene synthesis by using 2-step pcr

  • 1st PCR mixture
  1. 42.75 uL ddH2O7
  2. 1ul gfp 1-11 oligos (pooled) or gfp 12-22 pooled oligos
  3. 1.25 ul dNTP mix(10mM)
  4. 5 ul 10X pfu polymerase buffer with MgSO4
  5. 1 ul Pfu turbo hotstart polymerase
  • PCR conditions
  1. 95C for 2 min
  2. 95C for 30 sec
  3. 50C for 30 sec
  4. 72C for 1 min
  5. 72C for 2 min
  6. 4C unlimited time
  • results


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