Team:TU Darmstadt/Notebook/Methods/Dephosphorylation

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<p>Sie sind hier:&nbsp; <a href="index.php?id=23" >wiki</a> &rsaquo;&nbsp;<a href="index.php?id=32" >Notebook</a> &rsaquo;&nbsp;<a href="index.php?id=54" >Methods</a> &rsaquo;&nbsp;<span class="current"><a href="index.php?id=85" >Dephosphorylation</a></span></p>
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<!--TYPO3SEARCH_begin--><div id="c85" class="csc-default"><div><p><b>Short explanation:</b></p></div><div><p>Antarctic Phosphatase catalyzes the removal of 5' phosphate groups of DNA and RNA and thus prevents religation of cut vectors. It is used before ligation.</p></div><div></div><div><p><b>Procedure:</b></p></div><div><p>The reaction mix contains:</p></div><div><p>- up to 5 µg of digested DNA (no purification needed)</p></div><div><p>- 1/10 of reaction volume of 10x Antarctic Phosphatase Reaction Buffer</p></div><div><p>- 1 µL of Antarctic Phosphatase</p></div><div></div><div><p>- Incubate at 37° C for 30 minutes (3' ends) or 60 minutes (5' ends)</p></div><div><p>- Heat inactivate at 65° C for 15 minutes</p></div><div><p>- Continue with ligation</p></div></div><!--TYPO3SEARCH_end-->
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<!--TYPO3SEARCH_begin--><div id="c85" class="csc-default"><div class="csc-header csc-header-n1"><h1 class="csc-firstHeader">Dephosphorylation</h1></div><div><p><b>Short explanation:</b></p></div><div><p>Antarctic Phosphatase catalyzes the removal of 5' phosphate groups of DNA and RNA and thus prevents religation of cut vectors. It is used before ligation.</p></div><div></div><div><p><b>Procedure:</b></p></div><div><p>The reaction mix contains:</p></div><div><p>- up to 5 µg of digested DNA (no purification needed)</p></div><div><p>- 1/10 of reaction volume of 10x Antarctic Phosphatase Reaction Buffer</p></div><div><p>- 1 µL of Antarctic Phosphatase</p></div><div></div><div><p>- Incubate at 37° C for 30 minutes (3' ends) or 60 minutes (5' ends)</p></div><div><p>- Heat inactivate at 65° C for 15 minutes</p></div><div><p>- Continue with ligation</p></div></div><!--TYPO3SEARCH_end-->
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Latest revision as of 00:29, 18 October 2014

Home


Dephosphorylation

Short explanation:

Antarctic Phosphatase catalyzes the removal of 5' phosphate groups of DNA and RNA and thus prevents religation of cut vectors. It is used before ligation.

Procedure:

The reaction mix contains:

- up to 5 µg of digested DNA (no purification needed)

- 1/10 of reaction volume of 10x Antarctic Phosphatase Reaction Buffer

- 1 µL of Antarctic Phosphatase

- Incubate at 37° C for 30 minutes (3' ends) or 60 minutes (5' ends)

- Heat inactivate at 65° C for 15 minutes

- Continue with ligation

Retrieved from "http://2014.igem.org/Team:TU_Darmstadt/Notebook/Methods/Dephosphorylation"