User:Wesley J. Houston/Notebook/MAP/2013/03/08

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3/8/13

  • Ligation was repeated.
  • Attempted to grow more liquid cultures of ligated plasmids.
  • Attempted to confirm ligation by streaking the colonies on an agar plate.
  • All attempts proved unsuccessful.

> Ligations

  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
Ligation Plate results (lig : neg crtl) 03/08/13
1. KAH201 (S/P)/3403, 25 ng + KAH182 (X/P)/1675, 26 ng 0
2. KAH201 (S/P)/3403, 25 ng 0
  • No colonies were present in the ligation plate.


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