Fast DNA Digestion with Turbo Restriction Endonucleases

Standard Protocol and Conditions for Turbo Enzymes.

Properties of Turbo Restriction Endonucleases

Turbo enzymes have been tested for their ability to digest DNA within a short incubation time.
During testing, 1 μl of a Turbo enzyme was added to a reaction mixture containing 1 μg of DNA (phage or plasmid) and 1× universal ROSE buffer (for some enzymes, their own optimal buffer was used). After incubation for 10 or 15 minutes, complete digestion of the DNA substrate was observed.

Standard Turbo Digestion Reaction Protocol

Reaction volume: 20 μl

10× SE Buffer ROSE (or optimal buffer)

2 μl

DNA preparation

0,2–1 μg

BSA (10 mg/ml) (optional)*

0,2 μl

Sterile water

до 20 μl

+ 1 μl Turbo restriction endonuclease
/ or 1 μl of each of two Turbo enzymes for double digestion.

  1. Mix gently.
  2. Incubate at the optimal temperature for 10 or 15 minutes..

* Add to the reaction if BSA is required for maximal enzyme activity.

Notes

These enzymes should be used only in undiluted form.

For fast digestion, highly purified DNA preparations should be used, especially PCR fragments.
Complete digestion of some PCR products and supercoiled plasmid DNA may require longer than 10–15 minutes incubation due to their lower susceptibility to cleavage.