ezDNase™ Enzyme
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ezDNase™ Enzyme
Invitrogen™

ezDNase™ Enzyme

Invitrogen™ ezDNase™ Enzyme is a recombinant double-strand-specific DNase for the fast removal of contaminating genomic DNA from RNA preparations. ItRead more
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Catalog number 11766051
Price (USD)
288.00
Each
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Price (USD)
288.00
Each
Add to cart
Invitrogen™ ezDNase™ Enzyme is a recombinant double-strand-specific DNase for the fast removal of contaminating genomic DNA from RNA preparations. It cleaves phosphodiester bonds in double-stranded DNA to yield 2–8 bp oligonucleotides with 5’-phosphate and 3’-hydroxyl termini.

Features of ezDNase Enzyme include:
• Efficient and fast genomic DNA removal
• Highly specific—no impact on RNA, cDNA, or primers in RT reactions

ezDNase Enzyme's high specificity for double-stranded DNA enables efficient and fast genomic DNA removal without reduction in the quality or quantity of RNA or single-stranded DNA present in the reaction such as cDNA and primers. ezDNase Enzyme is heat-labile and so can be easily deactivated by heat treatment at moderate temperature (55°C). These features make ezDNase a superior choice for genomic DNA removal prior to reverse transcription reactions.

When used in combination with the SuperScript™ IV VILO™ Master Mix or other Invitrogen reverse transcription reagents, ezDNase Enzyme helps significantly reduce the possibility of cDNA synthesis being compromised by genomic DNA and reduce variation in RT-qPCR quantitation caused by RNA loss or damage during conventional DNase treatment.

For Research Use Only. Not for use in diagnostic procedures.
Specifications
Concentration10X
For Use With (Application)RT-qPCR
Shipping ConditionOn dry ice
EnzymeDNase
Compatible BufferReverse Transcription Reagents
Quantity50 μL
Product TypeEnzyme
Product LineezDNase™
Sample TypeRNA
FormLiquid
Unit SizeEach
Contents & Storage
• ezDNase Enzyme, 50 μL
• ezDNase Buffer (10X), 100 μL
• Nuclease-free water, 1.25 mL

Store at -5 to -30°C.

Frequently asked questions (FAQs)

How can I quantify the ssDNA in samples that contain both ssDNA and dsDNA, using the Qubit ssDNA Assay Kit (Cat. No. Q10212)?

Unfortunately, it is not possible to efficiently measure the ssDNA content in a sample that also contains dsDNA with the Qubit ssDNA Assay Kit (Cat. No. Q10212). The dye in the Qubit ssDNA Assay kit binds to both ssDNA and dsDNA.

It is possible to treat the sample with a DNase that only degrades dsDNA, such as theezDNase Enzyme (Cat. No. 11766051), and measure the ssDNA concentration afterward. However, if all dsDNA is not successfully digested, there is a risk of overestimating the ssDNA content.

The following article describes a PCR-based method to detect ssDNA in dsDNA samples:

Quantitative amplification of single-stranded DNA (QAOS) demonstrates that cdc13-1 mutants generate ssDNA in a telomere to centromere direction

Find additional tips, troubleshooting help, and resources within our Nucleic Acid Purification and Analysis Support Center.

Does the TaqMan Cells-to-CT Express Kit contain reagents for the removal of genomic DNA (gDNA)?

Yes, the TaqMan Cells-to-CT Express Kit includes Express ezDNase which can be added to the Express Lysis Solution for removal of gDNA during cell lysis. Express ezDNase is a double-strand specific, heat-labile DNase that will only digest gDNA, making it compatible with the downstream reverse transcription reaction. The enzyme is automatically inactivated during a heat kill step included in the reverse transcription program. To avoid the detection of gDNA, we recommend using a TaqMan Gene Expression Assay specifically designed to span an intron-exon boundary. Such assays are designated with an "_m1" suffix. More information can be found here.

Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Applications Support Center.

In your SuperScript IV RT protocols, there is no ezDNase inactivation step. Will active ezDNase affect RNA or the RT reaction?

The Invitrogen ezDNase Enzyme is a novel DNase that is highly specific for double-stranded DNA. It has no activity on single-stranded DNA in RT reactions (primers or probes), or on RNA. The enzyme is also thermolabile—it is inactivated quickly at temperatures typical for the SuperScript IV RT reaction (e.g., 50°C). The additional inactivation step is therefore not required in RT-qPCR applications.

Find additional tips, troubleshooting help, and resources within our PCR and cDNA Synthesis Support Center.

Which SuperScript IV RT format do you recommend for real-time PCR applications?

For RT-qPCR applications we recommend using the Invitrogen SuperScript IV VILO Master Mix (Cat. No. 11756050). The cDNA synthesis reaction setup with this master mix requires fewer pipetting steps and therefore reduces variation in the data. SuperScript IV RT, as a component of the master mix, offers the highest efficiency of cDNA synthesis step compared to competitors’ products.

Find additional tips, troubleshooting help, and resources within our PCR and cDNA Synthesis Support Center.

Are there any significant changes in the SuperScript IV RT protocol compared to the SuperScript III RT protocol?

The only change is that the incubation time for the reverse transcription reaction has been reduced from 50 minutes to 10 minutes. All the other parameters and steps are the same.

Find additional tips, troubleshooting help, and resources within our PCR and cDNA Synthesis Support Center.

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