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Team:Waterloo/Notebook/Protocols/Ligation
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(New page: <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> I...)
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(New page: <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> I...)
Newer edit →
Revision as of 18:10, 27 October 2008
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)
