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Endonucleases DNA-specific, dsDNase

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Double-Strand Specific dsDNase (dsDNase) is ideal for fast and effective removal of contaminating DNA from PCR master mixes.

Taq polymerases are commonly contaminated by bacterial DNA. This is a problem in PCR based bacterial typing and detection as it might cause false positive results. The unique properties of dsDNase make it suited for removal of contaminating DNA from PCR master mixes prior to addition of DNA template.

In figure 1, a PCR master mix was treated with different amounts of dsDNase before performing a qPCR to measure the contaminating bacterial DNA in the master mix. ArcticZymes dsDNase effectively removed contaminating DNA below known levels of the assay detection limits.

The dsDNase from Arctic shrimp (Pandalus borealis) is recombinantly produced in Pichia pastoris. It cleaves phosphodiester linkages in DNA to yield oligonucleotides with 5’-phosphate and 3’-hydroxyl termini.

The specific activity is estimated to be 30 times higher than that of bovine DNase I. In the presence of magnesium as only divalent cation and using oligos as a substrate, the activity towards dsDNA is 5000-fold higher than towards ssDNA.

The unique double strand-specificity allows specific degradation of dsDNA while leaving shorter ssDNA as primers and probes essentially intact. Easy inactivation by moderate heat (65°C) allows addition of DNA intended for analysis directly after removal of contaminating DNA.

Workflow – Decontamination of PCR master mixes

Recommended applications:

• Decontamination of PCR master mixes
• Removal of genomic DNA from RNA preparations

Key advantages with dsDNase:

• Double-strand DNA specific endonuclease
• High specific activity
• Can be heat-inactivated by moderate heat treatment (65°C for 15 minutes)
• Producing 5′-phospho-oligonucleotide products

Figures

Figure 1. The dsDNase effectively removes contaminated DNA

The dsDNase effectively removes contaminated DNA:

A PCR master mix was preincubated with various concentrations of dsDNase. After treatment, no DNA was amplified in non-template controls.

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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%

Double-Strand Specific dsDNase (dsDNase) is ideal for fast and effective removal of contaminating DNA from PCR master mixes.

Taq polymerases are commonly contaminated by bacterial DNA. This is a problem in PCR based bacterial typing and detection as it might cause false positive results. The unique properties of dsDNase make it suited for removal of contaminating DNA from PCR master mixes prior to addition of DNA template.

Product Description

    Product Detail

Applicable equipment:

It is recommended to use the Isothermal fluorescence detector developed by Amp-future, which is also suitable for fluorescence quantitative PCR apparatus with market known brands.

Kit Storage and Term of Validity

Storage term: stored at ≤-20℃,keep away from light, avoid heavy weight and repeated freezing and thawing.

Term of Validity: 14 months

Isothermal Nucleic Acid Principle Summary

This kit is based on a rapid nucleic acid amplification technology at room temperature and constant temperature: at room temperature and constant temperature (generally 39℃~42℃), with the help of auxiliary proteins and single-strand binding proteins,the recombinase and primers form a complex;Source search and combine the target homology domain, at this time,a D-loop region is formed at the homology position and strand exchange begins;along with the dissociation of the recombinase from the complex,the polymerase also binds to the 3′ end of the primer and begins chain extension. At the same time, relying on the function of exonuclease, adding specific molecular probes designed according to the template, and using fluorescence monitoring equipment can realize real-time monitoring of the amplification process of the target fragment.

Isothermal Nucleic Acid Product Features

1/ High sensitivity and specificity, short reaction time.

2/ The reagent form is freeze-dried, stable and easy to operate.

Technical Parameters:

Parameters	Details
Product Name	DNA Isothermal Amplification Kit EXO
Manufacturer	Amp-future
Storage Temperature	-20°C
Kit Components	Enzymes, Buffers ,Reagents
Packaging	48 Tests/box
Detection Limit	500-1000copies/µL
Shipping	ICE
Test Time	5-20mins

Isothermal Nucleic Acid Applications

Suitable for DNA isothermal rapid amplification kit(fluorescent type)

Primer: Require pair of nucleotide primers with the length of 25-35 bp.

Fluorescent Probe:Require the suitable length is 46-52nt.

DNA fluorescent kit reaction temperature is 39 to 42℃ and time is 5-20 minutes.

Notes

1/ Please avoid nucleic acid contamination and set blank control during reaction due to the high sensitivity of the kit.

2/ Please take out the required quantity of MIRA reaction units for the experiment, and put the rest under storage conditions when performing the experiment.

The NGS DNA Library Prep Kit (Ion Torrent platform) was developed for construction of high quality DNA libraries for next generation sequencing using Ion Torrent platform. The kit uses short double strand DNA fragments (blunt ends and/or sticky ends) as input DNA for NGS library construction, and is compatible with DNA fragments generated from both enzymatic methods (BioDynami DNA Fragmentation Enzyme Mix and DNA Fragmentation & A-tailing Enzyme Mix etc) and mechanical methods (sonication, nebulization etc.). Our unique technology increases library conversion efficiency and eliminates insert concatemer ligation. Library multiplexing up to 12 samples is possible.

NGS DNA Library Prep Kit Workflow

Kit features

• • • Simple procedure: Reactions for all three steps are in one tube
    • Fast protocol
      • Total protocol time is around 1 hour
      • Hands-on time is only ~5 minutes
    • Guaranteed high library conversion efficiency as compared to other kits
    • Input DNA amount: from 50 ng to 1 ug
    • No insert concatemer ligation

The NGS DNA Library Prep Kit (Ion Torrent platform) was developed for construction of high quality DNA libraries for next generation sequencing using Ion Torrent platform. The kit uses short double strand DNA fragments (blunt ends and/or sticky ends) as input DNA for NGS library construction, and is compatible with DNA fragments generated from both enzymatic methods (BioDynami DNA Fragmentation Enzyme Mix and DNA Fragmentation & A-tailing Enzyme Mix etc) and mechanical methods (sonication, nebulization etc.). Our unique technology increases library conversion efficiency and eliminates insert concatemer ligation. Library multiplexing up to 12 samples is possible.