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Protocol: Restriction digestion of Phytophthora infestans DNA for RG57 fingerprinting

  1. Determine the approximate concentration of your DNA sample by running a portion of the sample on a gel. We usually run 5 ul of a 50 ul DNA sample.
  1. Label a 1.5 ml microfuge tube for each digestion.
  2. Prepare a digestion mixture for each sample in the labelled 1.5 ml tube.
1-2 ug DNA x ul
spermadine (10 mg/ml) 2.0 ul
10X enzyme buffer 4.0 ul
Eco R1 enzyme 2.0 ul
RNase A (10 mg/ml) 1.0 ul
H2O to 40 ul
  1. Place the tubes in the 37° C incubator for at least 6 hr or overnight.
  1. Add running dye to each sample, load onto a 0.8 % agarose gel and run overnight at 30-40 volts.

infected tubers

infected tomatoes