User:Wesley J. Houston/Notebook/MAP/2013/03/13

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3/13/13

  • Attempted ligation following the same protocol outlined on 3/4/13.
  • Grew four liquid cultures of KAH182, following the same procedure (pick a colony and place into LB broth), in case this ligation attempt fails.
  1 2
Insert DNA (KAH182) 3.0 ---
Vector DNA (KAH201) 0.5 0.5
2x lgn buf (Roche) 5.0 5.0
T4 ligase (NEB) 1.0 1.0
dH2O 0.5 3.5
  10 μL 10 μL
  • Follow quick transformation protocol
  • Add 30 uL DH5α-Turbo to each ligation; plate on 100 μg/mL Amp agar


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