Team:Washington/COLONY PCR GREEN

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<p><strong>Colony  PCR with Green tag</strong><strong> </strong><br />
  (See <a href="https://2013.igem.org/Team:Washington/GENERAL_PCR_PROTOCOL">General PCR  Protocol</a> and <a href="https://2013.igem.org/Team:Washington/PCR_GOTAG">PCR GoTaq</a>) <br />
  <br />
  <strong>Master  mix (7ul):</strong> <br />
  1ul 10uM forward primer <br />
  1ul 10uM reverse Primer <br />
  5ul 2x Green taq (<a href="http://www.promega.com/%7E/media/Files/Resources/Protocols/Product%20Information%20Sheets/G/GoTaq%20DNA%20Polymerase%20M300.pdf">GoTaq</a>) <br />
  <br />
  <strong>Cell  water (3ul) &ldquo;colony water&rdquo;: </strong> <br />
  Pick one colony from the  plate and mix with 10ul of sterile diH2O or PBS to make 10ul of cell solution.  Save for inoculating overnights to miniprep. <br />
  If screening for successful  clones, the cell solution should be retained <br />
  Sterile glycerol can be  added to 50% final concentration, and cell solution can be frozen at -80oC for  use at a later date <br />
  *Make multiple cell waters  using all screened colonies and run multiple PCRs <br />
  <br />
  <strong>Reaction  Mixture:</strong> <br />
  Master mix (7ul) + Cell  solution (3ul) = 10ul total per tube <br />
  Use program  &quot;Colony&quot; &amp; change the extension time (1 min per kb) <br />
  <br />
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  <strong>PCR  Program: </strong> <br />
  &ldquo;Colony PCR&rdquo; <br />
  <br /></p>
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    <td width="76" valign="top"><p>1 Cycle </p></td>
    <td width="311" valign="top"><p>&nbsp;</p></td>
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    <td width="76" valign="top"><p>&nbsp;</p></td>
    <td width="311" valign="top"><p>94˚           3  minutes </p></td>
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    <td width="76" valign="top"><p>25 Cycles </p></td>
    <td width="311" valign="top"><p>&nbsp;</p></td>
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  <tr>
    <td width="76" valign="top"><p>&nbsp;</p></td>
    <td width="311" valign="top"><p>94˚           30  seconds <br />
55-60˚     30  seconds <br />
72˚           Amplicon  specific (~ 1 minute per kb) </p></td>
  </tr>
  <tr>
    <td width="76" valign="top"><p>1 Cycle </p></td>
    <td width="311" valign="top"><p>&nbsp;</p></td>
  </tr>
  <tr>
    <td width="76" valign="top"><p>&nbsp;</p></td>
    <td width="311" valign="top"><p>72˚          7  minutes <br />
4˚   &nbsp;   &nbsp;  &nbsp;     &nbsp;   &nbsp;     Forever</p></td>
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