User:Wesley J. Houston/Notebook/MAP/2013/03/18

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3/18/2013

  • Retrying ligation of KAH201(S/P)/3403 + KAH182(X/P)/1675
  • Concentrations: KAH201 = 48.1 ng/uL, KAH1182 = 6.3 ng/uL
    • Concentrations were determined on 2/18/13

> Ligations, following corrected version from 3/4/13, with 50 ng of vector instead of 25 ng.

  1 2
Insert DNA (KAH182) 8.0 ---
Vector DNA (KAH201) 1.0 1.0
2x lgn buf (Roche) 10.0 5.0
T4 ligase (NEB) 1.0 1.0
dH2O 0 3.0
  20 μL 10 μL
  • Follow quick transformation protocol
  • Add 30 uL DH5α-Turbo to each ligation; plate on 100 μg/mL Amp agar
Ligation Plate results (lig : neg crtl) 03/18/13
1. KAH201 (S/P)/3403, 50 ng + KAH182 (X/P)/1675, 50 ng 0
2. KAH201 (S/P)/3403, 50 ng 0