Salt-tolerant DNA & RNA Nuclease (heat-labile)
Salt-tolerant DNA & RNA Nuclease is a heat-labile non-specific endonuclease degrading double- and single-stranded DNA and RNA with optimum activity at high salt concentrations. It is active in a variety of buffers and can be easily inactivated by treatment with a reducing agent even at low temperatures. These features make Salt-tolerant DNA & RNA Nuclease particularly useful in the purification and removal of DNA and RNA after protein purification. The end-products after complete digestion consist mainly of oligos around 5 nucleotides. The enzyme is compatible with a wide range of buffers used in protein purification.
Main advantages of Salt-tolerant DNA & RNA Nuclease
- Non specific endonuclease
- Optimum activity at high salt concentration (0.5 M NaCl)
- Active at low temperatures (20% at 6ºC)
- Broad pH range
Recombinantly produced in Pichia pastoris.
Salt-tolerant DNA & RNA Nuclease is highly active in the temperature range 10–40°C. Optimal NaCl-concentration for activity is 0.5 M, working range is 0.25–1.0 M. Mg2 ; (>1 mM) is required for activity. Working pH range is 7.5–9.5, optimal pH is 9.0.
Minimum shelf life is 2 years at -20°C. Storage at 4°C is possible for at least 6 months. The enzyme tolerates multiple freeze-thaw cycles.
One unit is defined as the amount of Salt-tolerant DNA & RNA Nuclease that will cause a ΔA260 = 1 in 30 minutes at 37°C in a buffer consisting of 25 mM Tris-HCl, pH 8.5 (25°C), 5 mM MgCl2, 0.5 M NaCl and 50 μg/ml calf thymus DNA (D-1501, Sigma).
Inactivation of Salt-tolerant DNA & RNA Nuclease depends on the right combination of inactivation time, inactivation temperature (4–60°C) and concentration of the reducing agent (1–20 mM). If the reducing agent is completely removed after inactivation, trace amounts (<0.01 %) of Salt-tolerant DNA & RNA nuclease may reactivate. To avoid any reactivation, maintain a low concentration of reducing agent, 0.1–0.5 mM DTT or TCEP, alternatively use prolonged incubation times using 10–20 mM TCEP upon inactivation.
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