Team:TU Darmstadt/protocols/DNA Ligation

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<h2><font size="6" color="#F0F8FF" face="Arial regular"> DNA Ligation </font></h2>
<h2><font size="6" color="#F0F8FF" face="Arial regular"> DNA Ligation </font></h2>
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Latest revision as of 00:34, 5 October 2013







Lab book | Materials | Protocols

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

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