Team:Groningen/protocols/Ligation
From 2013.igem.org
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- | <li> | + | <li>MQ water</li> |
<li>1.5ml tubes</li> | <li>1.5ml tubes</li> | ||
<li>Ligation buffer</li> | <li>Ligation buffer</li> |
Revision as of 14:52, 27 July 2013
Ligation
Materials:
- MQ water
- 1.5ml tubes
- Ligation buffer
- Ligase
- Insert
- Vector
Reaction:
Component | 20μl reaction | Final concentration |
H2O | up to 20μl | |
10x Ligation buffer | 2μl | 1x |
Vector | 1μl | 20-100μg |
Insert | 1μl | 1:1 molar ratio |
T4 DNA Ligase 5U/μl | 0.2μl | 1U/μl |
All the reagents are added following the order listed in the table above.
After the reaction is ready mix the content of the tube and spin it down.
The tubes are incubated for at room temperature (~22°C) for 1h.
Reference: http://www.thermoscientificbio.com/uploadedFiles/Resources/fast-digestion-dna.pdf