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

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(gene synthesis by using 2-step pcr)
Current revision (08:24, 17 September 2010) (view source)
 
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|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Project name</span>
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|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> GENE SYNTHESIS</span>
|style="background-color: #F2F2F2" align="center"|<html><img src="/images/9/94/Report.png" border="0" /></html> [[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>[[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]]<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]]<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>}}
|style="background-color: #F2F2F2" align="center"|<html><img src="/images/9/94/Report.png" border="0" /></html> [[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>[[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]]<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]]<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>}}
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Current revision

GENE SYNTHESIS Main project page
Next entry

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

Image:1.pcr products.jpg


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