Protocol to cleanup DNA Glucosylation/restriction digest and Proteinase using Proteinase K (P8107)

  1. Mix 5-10 ug (30ug/ml final) genomic DNA, 80 uM UDP-glucose, 1xCutSmart Buffer™, and nuclease-free water to 155 ul volume.
  2. Add 30 U (3 ul) T4-β-glucosyltransferase (#M0357 , T4BGT).
  3. Incubate at 37 °C for 12 to 18 hrs.
  4. To 50 ul of T4BGT reaction, add 100 u (1 ul) restriction enzyme.
  5. Incubate at 37 °C 4-16 hrs, depending on restriction enzyme.
  6. Add 1 ul (800 U, ~20 ug) Proteinase K and incubate for 30 min at 40 °C.
  7. Inactivate Proteinase K by incubation for 10 min at 95 °C.