"
Team:TU Darmstadt/Notebook/Methods/DNA ligation
From 2014.igem.org
(Difference between revisions)
N.Hummel (Talk | contribs)
(Created page with "{{:Team:TU_Darmstadt/Template}} <html> <div id="contentWrap" class="container_24"> <div id="breadcrumbs" class="grid_24"> <p>Sie sind hier: <a href="http://20...")
Newer edit →
(Created page with "{{:Team:TU_Darmstadt/Template}} <html> <div id="contentWrap" class="container_24"> <div id="breadcrumbs" class="grid_24"> <p>Sie sind hier: <a href="http://20...")
Newer edit →
Revision as of 00:57, 16 October 2014
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: 20 µL batch
- T4 Ligase Buffer 2 µL
- T4 Ligase 1 µL
- digested vectors 25 ng (Molar ratio = Insert : backbone 3:1)
- add ddH2O to 20µL
- Incubate at 16°C overnight, alternatively incubate at room temperature for 30 min (results might be worse)
- Inactivate 20min at 80°C
