Haynes Lab:Notebook/Engineering PC-TFs/2014/05/12

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  • Using colony one of previously constructed CMV/MV9.
  • Restriction Digest
DNA(CMV/MV9) Concentration: 163ng/μL 5.0 μL
10X buffer 1.5 μL
XbaI 1.0 μL
dH2O 7.5 μL
Total 15 μL --> 37°C/ 15 min.
Mixture from Above Digest 15.0 μL
10X Phosphotase buffer 2.0 μL
Phosphotase 1.0 μL
dH2O 2.0 μL
Total 20 μL --> 37°C/ 10 min. 75°C for 2 min.

  • DNA Purification Kit standard protocol. Used 600μL binding buffer solution.
Plasmid OD260 OD260/280 ng/μL
1. Purified DNA 0.059 2.046 59.108
  • Eluted with 15μL (-1μL for concentration reading) H2O and put in Cameron box labeled Dephos XbaI CMV/MV9 5/12/14