Team:Brown-Stanford/Lab/Protocol/T4Ligation

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{{:Team:Brown-Stanford/Templates/Protocol}}
=='''T4 ligation'''==
=='''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])
(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])
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'''Reagents for 35µL reaction'''
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* 2.9 µL nuclease free water
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* 3.5 µL 10x T4 ligation buffer
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* 24 µL insert (BBa_K656013, 10 ng/µL)
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* 2.85 µL vector (pSCB3, 14.6ng/µL)
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* 1.75 µL T4 ligase
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'''Treatment'''
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# Add all reagents in order
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# Allow to incubate for 2 hours at room temperature
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# Denature ligase at 65˚C, store at 4˚ or -20˚

Latest revision as of 18:19, 28 September 2011

Brown-Stanford
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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

  1. Add all reagents in order
  2. Allow to incubate for 2 hours at room temperature
  3. Denature ligase at 65˚C, store at 4˚ or -20˚
Retrieved from "http://2011.igem.org/Team:Brown-Stanford/Lab/Protocol/T4Ligation"