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Cat. No.: HLDN-500 (for 500U)

Cat. No.: HLDN-5k (for 5000U)

Description

HL-dsDNase is a type of endonuclease that cleaves phosphodiester bonds in DNA, generating oligonucleotides with 5'-phosphate and 3'-hydroxyl ends. It specifically digests double-stranded DNA (dsDNA) without digesting single-stranded DNA, primers, probes, and RNA. HL-dsDNase is heat-sensitive and rapidly inactivated at 55°C. It is primarily used to quickly remove genomic DNA contamination from RNA samples before reverse transcription experiments. Compared to the traditional method of using DNase I to remove genomic DNA contamination, dsDNase does not require additional EDTA for inactivation, reducing RNA damage and saving experimental time while ensuring the accuracy of RNA quantification.

Unit definition

According to the Kunitz method, under conditions of pH 5.0 at 25°C, with excess high molecular weight DNA as the substrate, the amount of enzyme that causes an increase in absorbance at 260 nm of 0.001 per minute is defined as one unit (U) of activity.

Storage Buffer

25 mM Tris-HCl (pH 7.5, @25°C), 2.0 mM MgCl2, 10 mM NaCl, 0.01% (v/v) Triton X-100, 50% (v/v) glycerol.

Inhibition and Inactivation

• Inhibition conditions: Metal ions, EDTA, SDS, DTT, β-mercaptoethanol, high salt ion concentrations, etc., can inhibit the activity of dsDNase.
• Inactivation conditions: Incubation at 55°C for 5 minutes.

Quality Control

• Protein Purity: Assessed by SDS-PAGE and Coomassie brilliant blue staining, the enzyme purity is ≥90%.
• RNAse Activity Assay: Incubate the enzyme with RNA substrate at 37°C for 1 hour. Gel electrophoresis analysis shows no degradation of the RNA substrate.
• Functional Assay: The product is tested for the removal of genomic DNA from RNA samples and subsequent RT-qPCR amplification, demonstrating a removal rate of ≥99.9%. RNA quantity is not affected by dsDNase treatment.