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Team:TU Darmstadt/protocols/DNA Ligation
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Revision as of 12:38, 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
