Universal Flat Adapter for Heater-Shaker Module

endo-BCN-PEG3-acid is a click chemistry linker consisting of a BCN group with a terminal carboxylic acid. The terminal carboxylic acid can react with primary amine groups in the presence of activators (e.g. EDC, or HATU) to yield a amide bond. The BCN group can react with azide-tagged biomolecules. The hydrophilic PEG spacer increases solubility in aqueous media. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.

endo-BCN-PEG3-acid is a click chemistry linker consisting of a BCN group with a terminal carboxylic acid. The terminal carboxylic acid can react with primary amine groups in the presence of activators (e.g. EDC, or HATU) to yield a amide bond. The BCN group can react with azide-tagged biomolecules. The hydrophilic PEG spacer increases solubility in aqueous media. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.

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.