Tetherable DNA concentrations

I finished the gel extraction from the ligations of yesterday. So I decided to spare the 1ul required to measure the concentrations of the DNA for the nanodrop. I also measure the samples from the 3-piece ligations. Here are the resulting concentrations:

  • Some nomenclature:
    • T#pXX: T stands for tetherable; #indicates which adapter is in the final product (either 5’biotin or internal-biotin); pXX is the plasmid name for the adapter (either pRL or pALS)
  • T5pRL (~5.4kb)
    • 23.8ng/ul
    • 6.6nM
  • TIpRL (~5.4kb)
    • 9.3ng/ul
    • 2.6nM
  • T5pRL (~5.4kb, 3-piece result)
    • 14.7ng/ul
    • 4.1nM
  • TIpRL (~5.4kb, 3piece result)
    • 50.4ng/ul
    • 14nM
  • T5pALS (~8.4kb)
    • 23.6ng/ul
    • 4.3nM

Overall a much better yield than I expected, but I don’t completely trust the nanodrop so take these results to be whatever you want. In my experience I’ve never had good tethering efficiency with anything less than about 100pM DNA so I would recommend dilution of each of these about 1:10 for tethering purposes.