Double-strand specific DNases

Double-strand specific DNases (dsDNases) are unique double-strand specific endonucleases. As they do not digest ssDNA or RNA, they can be used to specifically remove dsDNA in the presence of other nucleic acids. The enzymes are heat-labile, which makes them ideal for applications where the DNase have to be inactivated. There are two versions of this enzyme available, dsDNase and HL-dsDNase.

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Available also in Glycerol-free and TRITON FREE formulations

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dsDNase and HL-dsDNase are now available

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Properties

Specificity towards double-stranded DNA
Nucleic acid specificity has been tested towards double- and single-stranded DNA and RNA oligonucleotides.

The specificity of dsDNase towards the substrate has been measured using 15-mer oligonucleotides with FAM at 5′ and DarkQuencher® 3′ (Eurogentec). The fluorescence is proportional to enzyme activity.

Assay conditions: 25 mM Tris pH 7.5, 5 mM MgCl2, and 2 μM oligonucleotide.

Relative activities
Substrate Relative activity
dsDNA 100%
ssDNA < 0.03%
dsRNA < 0.01 %
ssRNA < 0.01 %

From the data above we can conclude that the dsDNase is double-strand specific.

dsDNase

Heat inactivation
The dsDNase can be heat inactivated by heat treatment at 15 min at 65˚C, or 20 min at 60˚C. The enzyme requires 1 mM DTT and pH ≥ 8 for complete inactivation.

Specifications

  • Unit definition
    One Unit is defined as an increase in absorbance at 260 nm of 0.001 per minute, using 50 µg/ml high MW DNA in 100 mM Na-acetate pH 5.0 and 5 mM MgCl2.
  • Specific activity
    Ca. 400 000 Kunitz Units/mg.
  • Activity
    The dsDNase is highly active in a temperature range of 20-40°C. It needs at least 2.5 mM Mg for activity and has an optimal pH at 7.5.
  • Storage buffer
    20 mM Tris-HCl pH 7.5, 2 mM MgCl2, 10 mM NaCl, 0.01% (v/v) Triton X-100, 50% (v/v) glycerol.
  • Purity
    dsDNase is purified to apparent homogeneity.
  • Sample requirement
    Purification of nucleic acids prior dsDNase treatment.
  • Storage
    Minimum shelf life is 3 years at -20°C. Storage at 4°C is possible for at least 6 months. The enzyme also tolerates multiple freeze-thaw cycles.

HL-dsDNase

Heat inactivation
The HL-dsDNase can be heat inactivated by heat treatment at 5 min at 58˚C. The enzyme requires 1 mM DTT and pH ≥ 8 for complete inactivation.

Specifications

  • Unit definition
    One unit is defined as an increase in absorbance at 260 nm of 0.001 per minute, using 50 µg/ml of high MW DNA in 100 mM Na-acetate pH 5.0 and 5 mM MgCl2.
  • Specific activity
    Ca. 200 000 Kunitz Units/mg.
  • Activity
    The HL-dsDNase is highly active in a temperature range of 20-40°C. It needs at least 2.5 mM Mg for activity and has an optimal pH at 7.5.
  • Storage buffer
    20 mM Tris-HCl pH 7.5, 2 mM MgCl2, 10 mM NaCl, 0.01% (v/v) Triton X-100, 50% (v/v) glycerol.
  • Purity
    HL-dsDNase is purified to apparent homogeneity.
  • Sample requirement
    Purification of nucleic acids prior HL-dsDNase treatment.
  • Storage
    Minimum shelf life is 2 years at -20°C. The enzyme also tolerates multiple freeze-thaw cycles.
dsDNases brochure
Poster 1
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Publications

dsDNase Applications

  1. Adenosine and hyaluronan promote lung fibrosis and pulmonary hypertension in combined pulmonary fibrosis and emphysema.
    Collum SD, Molina JG, Hanmandlu A, Bi W, Pedroza M, Mertens TCJ, Wareing N, Wei W, Wilson C, Sun W, Rajadas J, Bollyky PL, Philip KM, Ren D, Thandavarayan RA, Bruckner BA, Xia Y, Blackburn MR, Karmouty-Quintana H.
    Disease Models & Mechanisms. 2019;12: dmm038711.

  2. The pause-initiation limit restricts transcription activation in human cells.
    Gressel S, Schwalb B, Cramer P.
    Nature Communications. 2019; 10: 3603.

  3. Metabolic effects of vitamin B12 on physiology, stress resistance, growth rate and biomass productivity of Cyanobacterium stanieri planktonic and biofilm cultures.
    Bohutskyi P, McClure RS, Hill EA, Nelson WC, Chrisler WB, Nuñez JR, Renslow RS, Charania MA, Lindemann SR, Beliaev AS.
    Algal Research. 2019; 42: 101580.

  4. Notum produced by Paneth cells attenuates regeneration of aged intestinal epithelium.
    Pentinmikko N, Iqbal S, Mana M, Andersson S, Cognetta III AB, Suciu RM, Roper J, Luopajärvi K, Markelin E, Gopalakrishnan S, Smolander OP, Naranjo S, Saarinen T, Juuti A, Pietiläinen K, Auvinen P, Ristimäki A, Gupta N, Tammela T, Jacks T, Sabatini DM, Cravatt BF, Yilmaz OH, Katajisto P.
    Nature. 2019; 571: 398-402.

  5. High Dietary Sugar Reshapes Sweet Taste to Promote Feeding Behavior in Drosophila melanogaster.
    May CE, Vaziri A, Lin YQ, Grushko O, Khabiri M, Wang QP, Holme KJ, Pletcher SD, Freddolino PL, Neely GG, Dus M.
    Cell Reports. 2019; 27 (6): 1675-1685.e7.

  6. First Complete Genome Sequences of Dengue Virus Serotype 2 Strains from the Solomon Islands and Vanuatu.
    Pyke AT, Cameron J, McMahon J, De Jong A, Burtonclay P.
    Microbiology Resource Announcements. 2019: 8(2):e01360-18.

  7. Illumina sequencing of clinical samples for virus detection in a public health laboratory.
    Huang B, Jenison A, Whiley D, McMahon J, Hewitson G, Grahma R, De Jong A, Warrilow D.
    Scientific Reports. 2019: 9: 5409.

  8. Environmental DNA metabarcoding of wild flowers reveals diverse communities of terrestrial arthropods.
    Thomsen PF, Sigsgaard EE.
    Ecololgy and Evolution. 2019; 9(4): 1665-1679.

  9. Rcs Phosphorelay Activation in Cardiolipin-Deficient Escherichia coli Reduces Biofilm Formation.
    Nepper JF, Lin YC, Weibel DB.
    Journal of Bacteriology. 2019; 201(9): e00804-18.

  10. Translocation of a Cell Surface Spliceosomal Complex Induces Alternative Splicing Events and Lymphoma Cell Necrosis.
    Tonapi SS, Pannu V, Duncan JE, Rosenow M, Helmstetter A, Magee D, Zhang Q, Tinder TT, Richards M, Halbert DD, Famulok M, Spetzler D, Miglarese MR, O'Neill HA, Mayer G.
    Cell Chemical Biology. 2019; 26(5): 756-764.e6.

  11. Impact of differential DNA methylation on transgene expression in cotton (Gossypium hirsutum L.) events generated by targeted sequence insertion.
    Verkest A, Bourout S, Debaveye J, Reynaert K, Saey B, Van den Brande I, K'Halluin K.
    Plant Biotechnol J. 2018; 17 (7): 1236-1247.

  12. Mechanical Genomic Studies Reveal the Role of D-Alanine Metabolism in Pseudomonas aeruginosa Cell Stiffness.
    Trivedi RR, Crooks JA, Auer GK, Pendry J, Foik IP, Siryaporn A, Abbott NL, Gitai Z, Weibel DB.
    mBio. 2018; 9(5): e01340-18.

  13. Angubindin-1 opens the blood–brain barrier in vivo for delivery of antisense oligonucleotide to the central nervous system.
    Zeniya S, Kuwahara H, Daizo K, Watari A, Kondoh M, Yoshida-Tanaka K, Kaburagi H, Asada K, Nagata T, Nagahama M, Yagi K, Yokota T.
    Journal of Controlled Release. 2018; 283: 126-134.

  14. Plerixafor enables safe, rapid, efficient mobilization of hematopoietic stem cells in sickle cell disease patients after exchange transfusion.
    Lagresle-Peyrou C, Lefrère F, Magrin E, Ribeil JA, Romano O, Weber L, Magnani A, Sadek H, Plantier C, Gabrion A, Ternaux B, Félix T, Couzin C, Stanislas A, Tréluyer JM, Lamhaut L, Joseph L, Delville M, Miccio A, André-Schmutz I, Cavazzana M.
    Haematologica. 2018; 103(5): 778.

  15. Buccal Mucosal Epithelial Cells Downregulate CTGF Expression in Buccal Submucosal Fibrosis Fibroblasts.
    Gottipamula S, Sundarrajan S, Moorthy A, Padmanabhan S, Srirdhar KN.
    J Maxillofac Oral Surg 2018; 17 (2), 254-259.

  16. Switching-Off Adora2b in Vascular Smooth Muscle Cells Halts the Development of Pulmonary Hypertension.
    Mertens TCJ, Hanmandlu A, Tu L, Phan C, Collum SD, Chen NY, Weng T, Davies J, Liu C, Eltzschig HK, Jyothula SSK, Rajagopal K, Xia Y, Guha A, Bruckner BA, Blackburn MR, Guignabert C, Karmouty-Quintana H.
    Front Physiol. 2018; 9, 555.

  17. Methyl-compound use and slow growth characterize microbial life in 2-km-depp subseafloor coal and shale beds.
    Trembath-Reichert E, Morono Y, Ijiri A, Hoshino T, Dawson KS, Inagaki F, Orphan VJ.
    Proceedings of the National Academy of Sciences. 2017; 114 (44) E9206-E9215.

  18. Zoantharian mitochondrial genomes contain unique complex group I introns and highly conserved intergenic regions.
    Chi SI, Johansen SD.
    Gene. 2017; 628:24-31.

  19. Seawater environmental DNA reflects seasonality of a coastal fish community.
    Sigsgaard EE, Nielsen IB, Carl H, Krag MA, Knudsen SW, Xing Y, Holm-Hansen TH, Møller PR, Thomsen PF.
    Marine Biology. 2017; 164:128.

  20. Microbial Typing by Machine Learned DNA Melt Signatures.
    Andini N, Wang B, Athamanolap P, Hardick J, Masek BJ, Thair S, Hu A, Avornu G, Peterson S, Cogill S, Rothman RE, Carroll KC, Gaydos CA, Wang JT, Batzoglou S, Yang S.
    Scientific Reports. 2017; 6 (7): 42097.

  21. Loss of CD73-mediated extracellular adenosine production exacerbates inflammation and abnormal alveolar development in newborn mice exposed to prolonged hyperoxia.
    Li H, Karmouty-Quintana H, Chen NY, Mills T, Molina J, Blackburn MR, Davies J.
    Pediatr Res. 2017; 82, 1039–1047.

  22. Inhibition of hyaluronan synthesis attenuates pulmonary hypertension associated with lung fibrosis.
    Collum SD, Chen NY, Hernandez AM, Hanmandlu A, Sweeney H, Mertens TCJ, Weng T, Luo F, Molina JG, Davies J, Horan IP, Morrell NW, Amione‐Guerra J, Al‐Jabbari O, Youker K Sun W, Rajadas J, Bollyky PL, Akkanti BH, Jyothula S, Sinha N, Guha A, Karmouty‐Quintana H.
    Br J Pharmacol. 2017; 174 (19), 3284-3301.

  23. A multiplexed amplicon approach for detecting gene fusions by next-generation sequencing.
    Beadling C, Wald AI, Warick A, Neff TL, Zhong S, Nikiforov YE, Corless LC, Nikiforova MN.
    The Journal of Molecular Diagnostics. 2016; 18 (2) 165 - 175.

  24. Extracellular adenosine levels are associated with the progression and exacerbation of pulmonary fibrosis.
    Luo F, Le NB, Mills T, Chen NY, Karmouty‐Quintana H, Molina JG, Davies J, Philip K, Volcik KA, Liu H, Xia Y, Eltzschig HK, Blackburn MR.
    FASEB J. 2016; 30 (2), 874-883.

  25. Palm-Sized Device for Point-of-Care Ebola Detection.
    Ahrberg CD, Manz A, Neužil P.
    Anal. Chem. 2016; 88 (9): 4803–4807.

  26. A contamination assessment of the CI carbonaceous meteorite Orgueil using a DNA-directed approach.
    Aerts, JW, Elsaesser A, Röling WFM and Ehrenfreund P.
    Meteoritics & Planetary Science. 2016; 51(5):920-931.

  27. Analysis of Ancient DNA in Microbial Ecology.
    Gorgé O, Bennett EA, Massilani D, Daligault J, Pruvost M, Geigl EM, Grange T.
    Microbial Environmental Genomics (MEG), Methods in Molecular Biology. 2016; 1399:289-315.

  28. Macrophage bone morphogenic protein receptor 2 depletion in idiopathic pulmonary fibrosis and Group III pulmonary hypertension.
    Chen NY, Collum SD, Luo F, Weng T, Le TT, Hernandez AM, Philip K, Molina JG, Garcia-Morales LJ, Cao Y, Ko TC, Amione-Guerra J, Al-Jabbari O, Bunge RR, Youker K, Bruckner BA, Hamid R, Davies J, Sinha N, Karmouty-Quintana H.
    Am J Physiol Lung Cell Mol Physiol. 2016; 311 (2); L238-54.

  29. The Influence of Tissue Procurement Procedures on RNA Integrity, Gene Expression, and Morphology in Porcine and Human Liver Tissue.
    Kap M, Sieuwerts AM, Kubista M, Oomen M, Arshad S, Riegman P.
    Biopreservation and Biobanking. 2015; 13(3): 200-206.

  30. Deletion of ADORA2B from myeloid cells dampens lung fibrosis and pulmonary hypertension.
    Karmouty-Quintana H, Philip K, Acero LF, Chen NY, Weng T, Molina JG, Luo F, Davies J, Le NB, Bunge I, Volcik KA, Le TT, Johnston RA, Xia Y, Eltzschig HK, Blackburn MR.
    FASEB J. 2015; 29 (1), 50-60.

  31. Identification of a microscopically selected microorganism in milk samples.
    N Bracke, Poucke MV, Baert B, Wynendaele E, Bels LD, Van Den Broeck W, Peelman L.
    Journal of Dairy Science. 2014; 97(2): 609–615.

  32. Double-stranded DNA in exosomes: a novel biomarker in cancer detection.
    Thakur BK, Zhang H, Becker A, Matei I, Huang Y, Costa-Silva B, Zheng Y, Hoshino A, Brazier H, Xiang J, Williams C, Rodriguez-Barrueco R, Silva JM, Zhang W, Hearn S, Elemento O, Paknejad N, Manova-Todorova K, Welte K, Bromberg J, Peinado H, Lyden D.
    Cell Research. 2014; 24:766–769.

  33. Association of markers of bacterial translocation with immune activation in decompensated cirrhosis.
    Mortensen C, Jensen JS, Hobolth L, Dam-Larsen S, Madsen BS, Andersen O, Møller S, Bendtsen F.
    Eur J Gastroenterol Hepatol. 2014; 26(12):1360-1366.
  34. Colonisation and interaction between S. epidermidis and S. aureus in the nose and throat of healthy adolescents.
    Fredheim EGA, Flægstad T, Askarian F, Klingenberg C.
    Eur J Clin Microbiol Infect Dis. 2014; 34, 123–129.

  35. Ancient DNA Analysis Reveals High Frequency of European Lactase Persistence Allele (T-13910) in Medieval Central Europe.
    Krüttli A, Bouwman A, Akgül G, Della Casa P, Rühli F, Warinner C.
    PLoS One. 2014; 9(1): e86251.doi: 10.1371/journal.pone.0086251.

  36. Adenosine promotes vascular barrier function in hyperoxic lung injury.
    Davies J, Karmouty-Quintana H, Le TT, Chen NY, Weng T, Luo F, Molina J, Moorthy B, Blackburn MR.
    Physiol Rep. 2014; 2 (9): e12155.

  37. Expansion of the CRISPR-Cas9 genome targeting space through the use of H1 promoter-expressed guide RNAs.
    Ranganathan V, Wahlin K, Maruotti J, Zacj DJ.
    Nat Commun. 2014; 5, 4516.

  38. No difference in portal and hepatic venous bacterial DNA in patients with cirrhosis undergoing transjugular intrahepatic portosystemic shunt insertion.
    Mortensen C, Karlsen S, Grønbæk H, Nielsen DT, Frevert S, Clemmesen JO, Møller S, Jensen JS, Bendtsen F.
    Liver Int. 2013; 33(9):1309–1315.

  39. Novel Sensitive Real-Time PCR for Quantification of Bacterial 16S rRNA Genes in Plasma of HIV-Infected Patients as a Marker for Microbial Translocation.
    Kramski M, Gaeguta AJ, Lichtfuss GF, Rajasuriar R, Crowe SM, French MA, Lewin SR, Center RJ, Purcell DF.
    J Clin Microbiol. 2011; 49(10): 3691–3693.

  40. An Efficient Multistrategy DNA Decontamination Procedure of PCR Reagents for Hypersensitive PCR Applications.
    Champlot S, Berthelot C, Pruvost M, Bennett EA, Grange T, Geigl EM.
    PLoS ONE. 2010; 5(9): e13042.

  41. Random DNA fragmentation allows detection of single-copy, single-exon alterations of copy number by oligonucleotide array CGH in clinical FFPE samples.
    Hostetter G, Kim SY, Savage S, Gooden GC, Barrett M, Zhang J, Alla L, Watanabe A, Einspahr J, Prasad A, Nickoloff BJ, Carpten J, Trent J, Alberts D, Bittner M.
    Nucleic Acids Research Advance Access. 2010; 38(2): e9.

  42. Bacterial communities of disease vectors sampled across time, space, and species.
    Jones RT, Knight R, Martin AP.
    The ISME Journal. 2010; 4(2):223-231.

  43. Experimental Murine Endometriosis Induces DNA Methylation and Altered Gene Expression in Eutopic Endometrium.
    Lee B, Du H, Taylor HS.
    Biology of Reproduction. 2009; 80(1): 79-85.

  44. Genetic characterization of the ABO blood group in Neandertals.
    Lalueza-Fox C, Gigli E, de la Rasilla M, Fortea J, Rosas A, Bertranpetit J, Krause J.
    BMC Evolutionary Biolog. 2008; 8:342.

  45. The dietary histone deacetylase inhibitor sulforaphane induces human β-defensin-2 in intestinal epithelial cells.
    Schwab M, Reynders V, Loitsch S, Steinhilber D, Schröder O, Stein J.
    Immunology. 2008; 125(2): 241–251.

  46. Quantification of reverse transcriptase in ALS and elimination of a novel retroviral candidate.
    McCormick AL, Brown RH Jr, Cudkowicz ME, Al-Chalabi A, Garson JA.
    Neurology. 2008; 70 (4): 278-283.

  47. The Derived FOXP2 Variant of Modern Humans Was Shared with Neandertals.
    Krause J, Lalueza-Fox C, Orlando L, Enard W, Green RE, Burbano HA, Hublin JJ, Hänni C, Fortea J, de la Rasilla M, Bertranpetit J, Rosas A, Pääbo S.
    Current Biology. 2007; 17 (21): 1908-12.

Descriptive Papers

  1. The Enzyme and the cDNA Sequence of a Thermolabile and Double-Strand Specific DNase from Northern Shrimps (Pandalus borealis).
    Nilsen I, Øverbø K, Havdalen LJ, Elde M, Gjellesvik DR , Lanes O.
    PLoS ONE. 2010; 5(4): e10295.