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Team:Groningen/Template/MODULE/Notebook/protocols/ligation
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| - | For ligation of DNA fragments 1U T4 DNA ligase was used. The amount of vector DNA that is used is 20 - 100 ng. The amount of insert in ng can be calculated using (ng of vector * kb of insert)/kb of vector * R with R being the molar ratio of the insert over the vector. The mixture is incubated for 1 hour at roomtemperature. After incubation, 5 μl of the mixture can be used for the transformation of 50 μl chemically competent cells or 1 μl for electrocompetent cells. | + | For ligation of DNA fragments 1U T4 DNA ligase was used. The amount of vector DNA that is used is 20 - 100 ng. The amount of insert in ng can be calculated using <i>(ng of vector * kb of insert)/kb of vector * R</i> with R being the molar ratio of the insert over the vector. The mixture is incubated for 1 hour at roomtemperature. After incubation, 5 μl of the mixture can be used for the transformation of 50 μl chemically competent cells or 1 μl for electrocompetent cells. |
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Latest revision as of 23:52, 17 October 2014
Ligation
For ligation of DNA fragments 1U T4 DNA ligase was used. The amount of vector DNA that is used is 20 - 100 ng. The amount of insert in ng can be calculated using (ng of vector * kb of insert)/kb of vector * R with R being the molar ratio of the insert over the vector. The mixture is incubated for 1 hour at roomtemperature. After incubation, 5 μl of the mixture can be used for the transformation of 50 μl chemically competent cells or 1 μl for electrocompetent cells.
