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Team:Waterloo/Notebook/Protocols/Ligation
From 2009.igem.org
(Difference between revisions)
| Line 16: | Line 16: | ||
- Vector DNA <br> | - Vector DNA <br> | ||
- Insert DNA <br> | - Insert DNA <br> | ||
| - | - | + | - 10X ligase buffer <br> |
- Ligase <br> | - Ligase <br> | ||
- MQ if needed <br> | - MQ if needed <br> | ||
3. Leave on the bench for 1 hour (or even overnight) <br> | 3. Leave on the bench for 1 hour (or even overnight) <br> | ||
Latest revision as of 02:56, 22 October 2009
Ligation (10µL recipe with 3 insert : 1 vector ratio)
Materials
MQ water
Ligase buffer
Ligase
Eppendorf tube
Vector DNA
Insert DNA
Instructions
1. Calculate ligation volumes. Total volume should be 10 µL and insert to vector ratio between 3:1 –5:1.
2. In one tube mix
- Vector DNA
- Insert DNA
- 10X ligase buffer
- Ligase
- MQ if needed
3. Leave on the bench for 1 hour (or even overnight)
