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Team:Groningen/protocols/Ligation
From 2013.igem.org
(Difference between revisions)
| Line 28: | Line 28: | ||
<li>MQ water</li> | <li>MQ water</li> | ||
<li>1.5ml tubes</li> | <li>1.5ml tubes</li> | ||
| - | <li>Ligation buffer</li> | + | <li>10x T4 Ligation buffer</li> |
| - | <li>Ligase</li> | + | <li>T4 Ligase</li> |
<li>Insert</li> | <li>Insert</li> | ||
<li>Vector</li> | <li>Vector</li> | ||
Revision as of 15:34, 27 July 2013
Ligation
Materials:
- MQ water
- 1.5ml tubes
- 10x T4 Ligation buffer
- T4 Ligase
- Insert
- Vector
Reaction:
| Component | 20µl | Final concentration |
|---|---|---|
| MilliQ water | up to 20µl | |
| 10 ligation buffer | 2µl | 1x |
| Vector | xµl | 20-100ng |
| Insert | xµl | 3:1 molar ratio over vector |
| T4 DNA ligase 5U/µl | 2µl | 0.5U/µl |
All the reagents are added following the order listed in the table above.
After finishing the reaction mixture, the content is mixed and incubated at room temperature (~22°C) for 20 min.
Reference: http://www.thermoscientificbio.com/uploadedFiles/Resources/fast-digestion-dna.pdf
iGEM 2013 Groningen
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