Features |
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Highly specific recombinant DNase for efficient removal of contaminating DNA |
• On-column DNA digestion – for time saving DNA removal during RNA preparation • DNA digestion in solution allows most efficient DNA removal for sensitive applications • No RNase activity detectable • RNA integrity (RIN) is not affected by rDNase treatment
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Technology |
Recombinant enzyme |
Source |
Recombinantly produced in Pichia pastoris without using any animal cells or other material derived from animals |
Format |
Lyophilized enzyme, separate Reaction Buffer
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RNase activity |
Not detectable* |
RNA integrity |
RNA integrity is unchanged by treatment with MACHEREY-NAGEL rDNAse under recommended conditions |
On-column DNA-removal reactions |
The amount of rDNase and Reaction Buffer is sufficient for**:
Mini spin columns: NucleoSpin® RNA: 50 preps NucleoSpin® miRNA***: 70 preps NucleoSpin® miRNA Plasma***: 140 preps NucleoSpin® RNA/Protein: 200 preps NucleoSpin® TriPrep: 200 preps NucleoSpin® RNA Blood: 125 preps NucleoSpin® RNA Plant: 50 preps NucleoSpin® totalRNA FFPE: 625 preps NucleoSpin® RNA Clean-up: 250 preps
Mini spin columns (XS design): NucleoSpin® RNA XS: 90 preps NucleoSpin® totalRNA FFPE XS: 240 preps NucleoSpin® RNA Clean-up XS: 90 preps
Midi spin columns: NucleoSpin® RNA Midi: 100 preps NucleoSpin® RNA Blood Midi: 50 preps
8-well format: NucleoSpin® 8 RNA: 240 preps NucleoSpin® 8 RNA Blood: 240 preps
96-well format: NucleoSpin® 96 RNA: 240 preps NucleoSpin® 96 RNA Blood: 240 preps
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Digestion in solution |
50 x 1 mL |
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Applications
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• On-column DNA-removal • Digestion in solution |
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* RNase activity is tested using a cleavable fluorescent labeled RNase substrate. No RNase activity is detectable after one hour incubation time. ** For correct use of the rDNase and Raction Buffer for rDNase in mentioned NucleoSpin RNA kits, please refer to the individual manual for detailed information. *** For NucleoSpin miRNA kits, dissolve rDNase in 1 mL Reaction Buffer for rDNase and transfer the rDNase solution back to the buffer bottle. Store and use rDNase as described in the individual manual.
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Principle/Procedure
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The rDNase Set is designed for use with NucleoSpin RNA Kits. It is suitable for time saving on-column DNA removal, as well as for most efficient DNA removal in the eluate. Furthermore, the rDNase Set is suitable for digestion of contaminating DNA within pre-purified RNA preparations (e.g. phenol based RNA preparations), i.e. for DNA digestion in solution.
Application data |
Samples of 100 µl crude RNA solution, contaminated with DNA, were treated with rDNase according to the protocol. Subsequently, the RNA was isolated using NucleoSpin RNA Clean-up XS. For the detection and determination of DNA and possible residual DNA, a qPCR reaction was performed before and after rDNAse treatment and clean-up. Results are shown in figure 1.
Analysis of RNA quality and quantity was performed using a Bioanalyzer and RNA 6000 Nano Reagent and chip (Agilent). Data of RNA integrity before and after rDNase digestion and clean-up are presented in figure 2. |
Efficient DNA removal from crude RNA extracts qPCR was performed before and after rDNase treatment of 3 samples (81 bp beta-Globin target; DyNamo Capillary SYBR® Green Kit (Finnzymes #F-420S/L)).
DNA contaminations are efficiently removed by rDNase digestion, resulting in Ct values, higher 35.
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Unchanged high RNA integrity after rDNase digestion RNA quality and quantity of the same sample were analyzed with an Agilent Bioanalyzer, Agilent RNA 6000 Nano chip, and Agilent RNA 6000 Nano reagent before and after rDNase digestion and clean-up.
MACHEREY-NAGEL rDNase shows high specificity. RNA integrity is unaffected by rDNase treatment and RIN remains unchanged, before and after rDNase digestion.
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*Increase of concentration is an effect of the RNA clean-up procedure with NucleoSpin RNA Clean-up XS
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Ordering information
Product |
Pack of
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Specification |
Reference |
rDNase Set |
1 set
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recombinant DNase and Reaction Buffer for rDNase, for 50 minipreparations of total RNA
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740963 |
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