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Team:Brown-Stanford/Lab/Protocol/T4Ligation
From 2011.igem.org
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T4 ligation
(Taken from [http://openwetware.org/wiki/DNA_Ligation openwetware.org]; this protocol was edited for amplification of [http://partsregistry.org/Part:BBa_K656013 Part BBa_K656013])
Reagents for 35µL reaction
- 2.9 µL nuclease free water
- 3.5 µL 10x T4 ligation buffer
- 24 µL insert (BBa_K656013, 10 ng/µL)
- 2.85 µL vector (pSCB3, 14.6ng/µL)
- 1.75 µL T4 ligase
Treatment
- Add all reagents in order
- Allow to incubate for 2 hours at room temperature
- Denature ligase at 65˚C, store at 4˚ or -20˚
