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rDNase Set
Features 
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

  
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

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: 800 preps
NucleoSpin® totalRNA FFPE XS: 1250 preps
NucleoSpin® RNA Clean-up XS: 800 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

Digestion in solution 50 x 1 mL

Applications

On-column DNA-removal
Digestion in solution

* 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.

  
     

Principle/Procedure

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. 

   

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.

  
*Increase of concentration is an effect of the RNA
  clean-up procedure with NucleoSpin RNA Clean-up XS

 
  
     

Ordering information

Product

Pack of

Specification Reference
rDNase Set

1 set

recombinant DNase and Reaction Buffer for rDNase,
for 50 minipreparations of total RNA

740963

For ordering click here

  
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