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Ligations
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Revision as of 16:07, 29 June 2010 by Timsterxox (Talk | contribs)
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
