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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> | <big><b><u>Ligation (10µL recipe with 5 insert : 1 vector ratio)</u></b></big> <br> | ||
Latest revision as of 22:54, 28 October 2008
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Ligation (10µL recipe with 5 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
- 10× ligase buffer
- Ligase
- MQ if needed
3. Leave on the bench for 1 hour (or even overnight)
