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Team:TU Delft/protocols/ligation
From 2010.igem.org
(Difference between revisions)
(New page: ==Ligation== Ligations (pasting plasmid DNA) were performed at the appropriate temperature with the appropriate buffer in the appropriate concentration, according to the supplier. React...) |
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|- | |- | ||
|DNA vector | |DNA vector | ||
| - | |x μL | + | |x μL |
|- | |- | ||
|T4 Ligation buffer (10×) | |T4 Ligation buffer (10×) | ||
| - | |2. | + | |2.0 μL (for 1×) |
|- | |- | ||
|T4 Ligase | |T4 Ligase | ||
| - | | | + | |1.0 μL |
|- | |- | ||
|H2O | |H2O | ||
| Line 22: | Line 22: | ||
|- | |- | ||
| | | | ||
| - | | | + | |20,0 μL (final contration ~20 ng/μL) |
|} | |} | ||
Transform circa half of the ligation mix. Incubate at 16 °C o/n | Transform circa half of the ligation mix. Incubate at 16 °C o/n | ||
Revision as of 19:18, 15 July 2010
Ligation
Ligations (pasting plasmid DNA) were performed at the appropriate temperature with the appropriate buffer in the appropriate concentration, according to the supplier.
Reaction for one sample:
| DNA insert | x μL |
| DNA vector | x μL |
| T4 Ligation buffer (10×) | 2.0 μL (for 1×) |
| T4 Ligase | 1.0 μL |
| H2O | x μL |
| 20,0 μL (final contration ~20 ng/μL) |
Transform circa half of the ligation mix. Incubate at 16 °C o/n
