User:Wesley J. Houston/Notebook/MAP/2013/03/25

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3/25/13

  • Prepped plasmids of KAH201 and determined concentration and purity.
  • Followed by digestion and gel electrophoresis, then purification.
  • However, concentrations were determined to be too low to complete a ligation.
  • More liquid cultures of KAH182 and KAH201 were prepped.
    • Colonies placed into LB broth overnight in shaking 37C incubator.
Sample 260/280 ng/μL
1. KAH201 (Plasmid) 1.795 54.054
2. KAH201 (Plasmid) 1.708 47.244


Reagent Volume   Results of the fourth electrophoresis attempt
30 μL/lane, 1% agarose; Ladder
DNA (plasmid) 25 μL
10x buffer 3.0
enzyme 1 (Pst1) 1.0
enzyme 2 (Xba1 or Spe1) 1.0
dH2O 0.0 μL
  30 μL --> 37°C/ ~15 min.

> Measure conc.'s of gel purified fragments

Sample 260/280 ng/μL
1. KAH201 (S/P) 0.727 2.54
2. KAH182 (X/P) 0.789 1.596
  • Due to the low concentrations, I made more liquid cultures of KAH182 and KAH201.
    • Followed the established protocol for making liquid cultures: pick one colony and place into LB broth overnight in shaking 37C incubator.