Team:Groningen/protocols/Ligation
From 2013.igem.org
(Difference between revisions)
Line 64: | Line 64: | ||
<br>All the reagents are added following the order listed in the table above. | <br>All the reagents are added following the order listed in the table above. | ||
<br>After the reaction is ready mix the content of the tube and spin it down. | <br>After the reaction is ready mix the content of the tube and spin it down. | ||
- | <br>The tubes are incubated for | + | <br>The tubes are incubated for at room temperature (~22°C) for 1h. |
<br> | <br> | ||
<br>Reference: http://www.thermoscientificbio.com/uploadedFiles/Resources/fast-digestion-dna.pdf | <br>Reference: http://www.thermoscientificbio.com/uploadedFiles/Resources/fast-digestion-dna.pdf |
Revision as of 10:08, 26 July 2013
Ligation
Materials:
- H2O
- 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 | 0.2μl | 5U/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