Team:TU Darmstadt/protocols/DNA Ligation

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<img alt="Problem" src="/wiki/images/6/66/Darmstadt_green_Problem.jpg" width="100" height="30"></a>
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<a href="https://2013.igem.org/Team:TU_Darmstadt/labbook">
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<!-- DNA Ligation -->
<!-- DNA Ligation -->

Revision as of 23:45, 4 October 2013





DNA Ligation

Short explanation
 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.

Procedure
 Ligation (20 µL batch)
T4 Ligase Buffer 2 µL
T4 Ligase 1 µL
digested vector 25 ng
Insert 1:3 molar ratio compared to vector
add to 20µL with ddH2O

Incubate at 16°C over night
inaktivate 20min by 65°C

Retrieved from "http://2013.igem.org/Team:TU_Darmstadt/protocols/DNA_Ligation"