Team:Waterloo/Notebook/Protocols/Ligation

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<big><b><u>Ligation (10µL recipe with 5 insert : 1 vector ratio)</u></b></big> <br>

<b>Materials </b><br>
MQ water <br>
Ligase buffer<br> 
Ligase <br>
Eppendorf tube<br>
Vector DNA<br>
Insert DNA<br>

<b>Instructions </b><br>
1. Calculate ligation volumes. Total volume should be 10 µL and insert to vector ratio between 3:1 –5:1. <br>
2. In one tube mix<br>
- Vector DNA <br>
- Insert DNA <br>
- 10× ligase buffer <br>
- Ligase <br>
- MQ if needed <br>
3. Leave on the bench for 1 hour (or even overnight) <br>