Haynes Lab:Notebook/Engineering PC-TFs/2012/02/16
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Ligation/Assembly
Measure concentration of each fragment:
Results:
Assembly Scheme: Number beside vector and insert is number of base pairs
> Warm five 100 μg/mL Amp agar plates at 37 °C > Thaw fresh tube of DH5α Turbo cells on ice > Resuspend invisible DNA pellet (stock DNA) in 10 μL dH2O > Add 0.5 μL stock DNA to 10 μL dH2O in 0.5 mL tubes > Add 30 μL of DH5α Turbo cells to DNA + dH2O --> Include #5 tube, water only, no DNA (negative control) > Incubate cells + DNA on ice for 5 min. > Label pre-warmed plates > Transfer cells + DNA onto agar > Add 10 - 15 sterile glass beads, shake, discard beads > Incubate plates at 37 °C overnight
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