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Duplex DNase

Catalog # Concentration Size List Price Quantity Your Price
M7635L 750 units $928.00
$835.20
M7635S 150 units $232.00
$208.80
Catalog # Size List Price Your Price
M7635L 750 units $928.00
$835.20
M7635S 150 units $232.00
$208.80
Catalog #
Qty:
 
*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca
M7635 Mechanism
  • DNA-specific endonuclease
  • Specifically degrades dsDNA in the presence of ssDNA
  • Cleaves DNA strand of DNA:RNA hybrid duplex
  • Products are short oligos or ssRNA (if cleaving the DNA strand of DNA:RNA hybrid)

    Duplex DNase is ideal for:
  • Removal of contaminating gDNA prior to RT-qPCR
  • Removal of contaminating DNA from RNA or protein preps
  • PCR decontamination
  • Consider DNase I (RNase-free) or DNaseI-XT for IVT applications

Duplex DNase is an engineered double-strand-specific DNA endonuclease that preferentially degrades double-stranded DNA (dsDNA) over single-stranded DNA (ssDNA) or RNA. It will also cleave the DNA strand of a DNA:RNA hybrid duplex.

Figure 1: Duplex DNase preferentially degrades dsDNA over ssDNA

M7635 Degradation

Activity on a dsDNA substrate (35 nt dsDNA hairpin) or ssDNA substrate (15 nt ssDNA) was measured by monitoring the increase in fluorescence when the substrate is degraded by Duplex DNase. FLUOR = Fluorophore; QUENCH = Quencher


Figure 2: Duplex DNase is more specific for dsDNA than DNase I


M7635 dsDNA vs ssDNA

Activity of Duplex DNase (NEB #M7635) or DNase I (NEB #M0303) on a dsDNA substrate (35 nt dsDNA hairpin) or ssDNA substrate (15 nt ssDNA) was measured by monitoring the increase in fluorescence when the substrate is degraded by either enzyme in their respective reaction buffers at 30°C. Duplex DNase is > 5-fold more specific than DNase I is for dsDNA. The ratio of activity on double-stranded vs. single-stranded substrates demonstrates the increased specificity that Duplex DNase shows for dsDNA over ssDNA as compared to DNase I.


Figure 3: Duplex DNase degrades the DNA strand of a DNA:RNA hybrid

M7635 Hybrid Degradation

An in vitro transcription reaction (50 μl) was treated with 2 U DNase I-XT for 15 minutes at 37°C to digest template DNA. The RNA was purified using the Monarch® RNA Cleanup Kit (500 µg, NEB #T2050) and eluted in nuclease-free water (50 μl). Varying amounts (from 106-1012 copies) of Cluc template RNA were mixed with a 5´ fluorescently-labeled/3´-quenched 22 nt complementary DNA probe (0.2 µM final concentration, Tm 61.8°C) in NEBuffer™ r1.1. Duplex DNase (2U) was added, the reaction incubated at 58°C, and the relative fluorescence was monitored every 10 seconds in a Bio-Rad® CFX Touch™ qPCR instrument. The increase in fluorescence observed over time (which is not observed in the non-template control) demonstrates the ability of Duplex DNase to cleave the fluorescently labeled DNA strand of a DNA:RNA hybrid (probe:Cluc RNA). Moreover, the cleavage of the DNA probe when hybridized to the template RNA increases over time and is dose-dependent on the amount of Cluc template RNA present. FLUOR = Fluorophore; QUENCH = Quencher


Figure 4: Duplex DNase preferentially degrades dsDNA in a mixture of dsDNA and ssDNA over a wide range of temperatures

M7735 Degradation Analysis

A mixture of 20 pmol of dsDNA (60 bp oligonucleotide) and 20 pmol of ssDNA (50 nt oligonucleotide) was incubated with 2 Units of Duplex DNase for 1 minute at 25°C, 37°C, 50°C or 65°C and then run on a 4% E-gel™ EX gel (Thermo Fisher Scientific®) stained with SYBR® Gold. Lane 1: dsDNA PCR Marker (NEB #N3234), Lane 2: ssDNA 50 nt, Lane 3: dsDNA 60 bp, Lanes 4–7: 25°C, 37°C, 50°C, 65°C. Duplex DNase displays increased activity and preference for double stranded DNA with increasing temperatures. FLUOR = Fluorophore


Product Source
A His-tagged engineered Duplex DNase expressed in Pichia pastoris.
Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  Duplex DNase M7635SVIAL -20 1 x 0.075 ml 2,000 units/ml
  NEBuffer™ r1.1 B6001SVIAL -20 1 x 1.25 ml 10 X
  Duplex DNase M7635LVIAL -20 1 x 0.375 ml 2,000 units/ml
  NEBuffer™ r1.1 B6001SVIAL -20 2 x 1.25 ml 10 X
An exception occurred during the operation, making the result invalid. Check InnerException for exception details.

Companion Products
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Certificate of Analysis
The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality controls for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]
Safety Data Sheets
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