Team:TU Darmstadt/Notebook/Methods/DNA ligation

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<!--TYPO3SEARCH_begin--><div id="c86" class="csc-default"><div class="csc-header csc-header-n1"><h1 class="csc-firstHeader">DNA ligation</h1></div><div><p><b>Short explanation:</b></p></div><div><p>DNA ligation is necessary to assemble digested DNA parts into a vector. The cut ends generated by restriciton enzymes are put together by DNA ligase.</p></div><div></div><div><p><b>Procedure:&nbsp;</b>20 µL batch</p></div><div><p>- T4 Ligase Buffer 2 µL
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<!--TYPO3SEARCH_begin--><div id="c86" class="csc-default"><div class="csc-header csc-header-n1"><h1 class="csc-firstHeader">DNA Ligation</h1></div><div><p><b>Short explanation:</b></p></div><div><p>DNA ligation is necessary to assemble digested DNA parts into a vector. The cut ends generated by restriciton enzymes are put together by DNA ligase.</p></div><div></div><div><p><b>Procedure:&nbsp;</b>20 µL batch</p></div><div><p>- T4 Ligase Buffer 2 µL
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<p>- T4 Ligase 1 µL</p></div><div><p>- digested vectors 25 ng (Molar ratio = Insert : backbone 3:1)</p></div><div><p>- add ddH2O to 20µL</p></div><div><p>- Incubate at 16°C overnight, alternatively incubate at room temperature for 30 min (results might be worse)</p></div><div><p>- Inactivate 20min at 80°C</p></div></div><!--TYPO3SEARCH_end-->
<p>- T4 Ligase 1 µL</p></div><div><p>- digested vectors 25 ng (Molar ratio = Insert : backbone 3:1)</p></div><div><p>- add ddH2O to 20µL</p></div><div><p>- Incubate at 16°C overnight, alternatively incubate at room temperature for 30 min (results might be worse)</p></div><div><p>- Inactivate 20min at 80°C</p></div></div><!--TYPO3SEARCH_end-->

Latest revision as of 00:31, 18 October 2014

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