Team:Warsaw/Calendar-Main/26 June 2008

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Preparation of pMPMT5-AID+AIDT7 construct
1. Transformation of E. coli TOP10 strain with ligation from previous day
2. Plating transformants on LB + tetracycline

Polymerase Chain Ligation (PCL)

ProductTemplatesPrimersProduct length
Alpha-A fusionAlpha-linker + linker-AAlphaL+SacI + AP+NotI 1500 bp
Omega-A fusionOmega-linker + linker-AOmegaL+SacI + AP+NotI 1440 bp

PCL program for fusion A with alpha and omega

TemperatureTime
94°C4:00
94°C0:3028 cycles
48°C-62°C0:45
72°C2:30
72°C10:00
4°Cinfinite

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