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Ligations
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Timsterxox (Talk | contribs) (New page: '''Reagents''' *T4 DNA ligase *10x T4 DNA Ligase Buffer *Deionized, sterile H2O *Purified, linearized vector (likely in H2O or EB) *Purified, linearized insert (likely in H2O or EB) '''E...) |
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*Add (8.5 - vector and insert volume)μl ddH2O | *Add (8.5 - vector and insert volume)μl ddH2O | ||
*0.5 μL T4 Ligase | *0.5 μL T4 Ligase | ||
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| + | '''Calculating Insert Amount''' | ||
| + | [[Image: insert.jpg|center|150px]] | ||
Revision as of 16:05, 29 June 2010
Reagents
- T4 DNA ligase
- 10x T4 DNA Ligase Buffer
- Deionized, sterile H2O
- Purified, linearized vector (likely in H2O or EB)
- Purified, linearized insert (likely in H2O or EB)
Equipment
- Vortex
Procedure
10μL Ligation Mix
Note: Larger ligation mixes are also commonly used
- 1.0 μL 10X T4 ligase buffer
- 6:1 molar ratio of insert to vector (~10ng vector)
- Add (8.5 - vector and insert volume)μl ddH2O
- 0.5 μL T4 Ligase
Calculating Insert Amount
