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Quick Ligation Reaction Buffer
- Combine 50 ng of vector with a 3-fold molar excess of insert. Use NEBioCalculator to calculate molar ratios. Adjust volume to 10 μl with dH2O.
- Add 10 μl of 2X Quick Ligation Buffer and mix.
- Add 1μl of Quick T4 DNA Ligase and mix thoroughly.
- Centrifuge briefly and incubate at room temperature (25°C) for 5 minutes.
- Chill on ice, then transform or store at -20°C.
- Do not heat inactivate. Heat inactivation dramatically reduces transformation efficiency.