Team:Groningen/protocols/Ligation
From 2013.igem.org
(Difference between revisions)
Line 22: | Line 22: | ||
<div class="mainContent"> | <div class="mainContent"> | ||
- | + | <h1>Ligation</h1> | |
+ | |||
+ | <h3>Materials:</h3> | ||
+ | <ul type="square"> | ||
+ | <li>H<sub>2</sub>O</li> | ||
+ | <li>1.5ml tubes</li> | ||
+ | <li>Ligation buffer</li> | ||
+ | <li>Ligase</li> | ||
+ | <li>Insert</li> | ||
+ | <li>Vector</li> | ||
+ | </ul> | ||
+ | |||
+ | <h3>Reaction:</h3> | ||
+ | <table border = "1"> | ||
+ | <tr><td>Component</td> | ||
+ | <td>30μl reaction</td> | ||
+ | <td>Final concentration</td> | ||
+ | </tr> | ||
+ | <tr><td>H<sub>2</sub>O</td> | ||
+ | <td>up to 30μl</td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr><td>Buffer 10x</td> | ||
+ | <td>3μl</td> | ||
+ | <td>1x</td> | ||
+ | </tr> | ||
+ | <tr><td>DNA</td> | ||
+ | <td>4μl</td> | ||
+ | <td>up to 1μg</td> | ||
+ | </tr> | ||
+ | <tr><td>Enzyme/s</td> | ||
+ | <td>0.5μl</td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | |||
+ | <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>The tubes are incubated for 1h at 37°C. | ||
+ | <br> | ||
+ | <br>Reference: http://www.thermoscientificbio.com/uploadedFiles/Resources/fast-digestion-dna.pdf | ||
</div> | </div> |
Revision as of 09:56, 26 July 2013
Ligation
Materials:
- H2O
- 1.5ml tubes
- Ligation buffer
- Ligase
- Insert
- Vector
Reaction:
Component | 30μl reaction | Final concentration |
H2O | up to 30μl | |
Buffer 10x | 3μl | 1x |
DNA | 4μl | up to 1μg |
Enzyme/s | 0.5μl |
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 1h at 37°C.
Reference: http://www.thermoscientificbio.com/uploadedFiles/Resources/fast-digestion-dna.pdf