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Nucleic acid purification

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Product overview

Magnetic-bead-based isolation of viral RNA/DNA and bacterial DNA from veterinary samples

The NucleoMag VET kit is designed for the isolation of viral RNA/DNA and bacterial DNA from common veterinary samples such as whole blood, serum/plasma, feces, ear notches, or swabs using manual or automated processing. The procedure is based on the reversible adsorption of nucleic acids to paramagnetic beads. Sample lysis is achieved by incubation with a lysis buffer containing chaotropic ions and supplemented by Proteinase K digestion. Nucleic acids are bound to paramagnetic NucleoMag B-Beads in the presence of Binding Buffer VEB. After magnetic separation, the paramagnetic beads are subjected to successive wash steps to remove contaminants and salts, and highly pure RNA/DNA are eluted in low-salt Elution Buffer VEL or water.

  • One kit for common veterinary samples
  • High sensitivity, even with low viral titers
  • Robust, one-tube procedure minimizes risk of cross-contamination
  • Kit format allows for manual or automated processing on common liquid handling platforms

At-a-glance  

NucleoMag VET
Technology Magnetic bead technology
Format Highly reactive superparamagnetic beads
Processing Manual or automated
Starting material* <200 µl whole blood, serum, plasma
<30 mg tissue (e.g., ear notches)
<200 µl feces
<200 µl swab wash solution
Fragment size 300 bp–approximately 50 kb
Elution volume 50–100 µl
Preparation time 45–120 min/96 preps (45 min for KingFisher)
Binding capacity 0.4 μg/μl beads

*For all sample types, start with a maxiumum of 200 µl liquid/homogenized sample.

NucleoMag principle  

The NucleoMag family of products is based on magnetic bead technology. Nucleic acids are bound to the magnetic beads under chaotropic salt conditions, contaminants are washed away with ethanolic wash buffers, and pure RNA and DNA are eluted from the beads using elution buffer. 

NucleoMag bind

Figure 1. NucleoMag magnetic bead technology. NucleoMag beads and binding buffer are added to the sample (left). Beads bound to DNA/RNA are held in place in the well by the magnet, and contaminants are washed away (middle). DNA/RNA is eluted while the beads are held in place by the magnet (right).

Procedure  

NucleoMag SEP

Figure 2. Overview of NucleoMag VET protocol for manual processing of 96 samples using NucleoMag SEP.

Example data  

Graph: detection of viral DNA from animal tissue samples

Figure 3. Sensitive detection of viral DNA from animal tissue samples. Viral DNA was purified from PCV2-positive animal tissue samples (n = 22, 20 mg each) using NucleoMag VET and a kit from Competitor Q, respectively, and analyzed by qPCR. Of the 22 samples tested, 20 were found to be positive for PCV2 using NucleoMag VET, whereas only 14 were found to be positive using the kit from Competitor Q. The graph depicts mean CT values for amplification of the PCV2 sequence from viral DNA purified with either kit.

Graph: Detection of viral RNA and DNA from animal blood samples

Figure 4. Sensitive detection of viral RNA and DNA from animal blood samples. Purified Viral RNA and DNA obtained from 200 µl blood serum samples of PRRSV- and PCV2-positive animals using NucleoMag VET was diluted 10-fold and analyzed by qPCR. For both RNA and DNA (blue and gray bars, respectively), CT values correlate with dilution factor. Sequences corresponding to PRRSV and PCV2 were detectable across the range of dilutions tested.

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Isolation of viral RNA and DNA and bacterial DNA from cell-free body fluids,homogenized tissue sample suspensions, stool sample suspensions, and swab washes

RNA and DNA isolation from pathogens

The NucleoMag Pathogen kit is designed for the rapid manual and automated small-scale isolation of viral RNA and DNA and bacterial DNA from cell-free body fluids such as serum or plasma, blood, homogenized tissue sample suspensions, stool sample suspensions, and swab washes.

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