Sentelab:Notebook/gene synthesis/2010/09/17

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(Autocreate 2010/09/17 Entry for Sentelab:Notebook/gene_synthesis)
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==Entry title==
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==gene synthesis by using 2-step pcr==
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* Insert content here...
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* 1st PCR mixture
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#42.75 uL ddH2O7
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#1ul gfp 1-11 oligos (pooled) or gfp 12-22 pooled oligos
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#1.25 ul dNTP mix(10mM)
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#5 ul 10X pfu polymerase buffer with MgSO4
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#1 ul Pfu turbo hotstart polymerase
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*PCR conditions
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#95C for 2 min
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#95C for 30 sec
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#50C for 30 sec
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#72C for 1 min
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#72C for 2 min
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#4C unlimited time

Revision as of 08:14, 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



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