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Home › Learning centers › Nucleic acid purification › Genomic DNA purification › Blood and plasma › NucleoMag cfDNA

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Isolate circulating DNA from plasma NucleoSnap cfDNA
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Product overview

High-throughput isolation of cell-free DNA from plasma

The NucleoMag cfDNA kit is designed for the efficient isolation of circulating cell-free DNA (cfDNA) from human blood plasma. Fragmented DNA as small as 50 bp can be purified at high yield due to a reversible adsorption of nucleic acids to paramagnetic beads under specialized buffer conditions. The yield can vary depending on the sample and is typically in the range of 0.1 to 100 ng DNA per ml of plasma. The purification procedure takes less than an hour for 24 preps.

  • Consistent cfDNA recovery from 1–10 ml of plasma
  • Efficient purification of fragmented DNA as small as 50 bp
  • No PCR inhibition
  • Manual or high-throughput automated isolation of circulating cell-free DNA from human plasma using EDTA or Cell-Free DNA BCT tubes
  • Typical downstream applications: NGS, qPCR, ddPCR
At-a-glance Higher yield Highly consistent Expected yield Protocol schematic

At-a-glance  

Technology Magnetic bead technology
Format Highly reactive superparamagnetic beads
Processing Manual or automated
Starting material 1–10 ml human EDTA/Cell-Free DNA BCT plasma
Fragment size >50 bp
Typical yield Depends on sample source, storage, and quality
A260/A280 1.6–1.9
Elution volume 50-200 µl
Preparation time 55 min/24 preps (2 ml; excluding lysis)*
Binding capacity 0.3 µg/µl beads

 
* Using a KingFisher Flex

Higher yield  

Higher yields from low-abundance cfDNA samples.

Higher yields than a leading competitor from low-abundance cfDNA samples. Total cfDNA was purified from 2 ml of human EDTA plasma derived from four challenging donor samples known to have a low abundance of cfDNA (<10 ng cfDNA/ml plasma). Purification using the NucleoMag cfDNA kit resulted in higher and more consistent yields of total cfDNA compared to the competitor. The final total DNA recovery was quantified using the Qubit dsDNA High Sensitivity kit on a Qubit fluorometer (Thermo Fisher Scientific).

Highly consistent  

Consistent recovery of cfDNA regardless of plasma volume.

Consistent recovery of cell-free DNA regardless of plasma volume. cfDNA from human EDTA plasma isolated using the NucleoMag cfDNA kit shows clear bands when analyzed via capillary gel electrophoresis (using the Agilent Bioanalyzer 2100 system with the High Sensitivity DNA kit). cfDNA is clearly detectable, even when it is purified from small volumes of plasma.

Expected yield  

DNA yield ranges from 0.1 to 100 ng DNA per ml plasma.

DNA yield strongly depends on individual samples, but is typically in the range of 0.1 to 100 ng DNA per ml of plasma. The NucleoMag cfDNA kit was used to isolate cfDNA from human EDTA plasma samples ranging from 2–10 ml in volume. Efficient purification of cfDNA was indicated by the characteristic peak at approximately 170 bp (using the Agilent Bioanalyzer 2100 system and the High Sensitivity DNA kit). The total yield was determined by PicoGreen quantification assay using the Quant-iT PicoGreen kit (Thermo Fisher Scientific) on a Synergy HT Multi-detection microplate reader (Biotek). The consistent correlation of cfDNA amount and sample volume demonstrated reliable performance independent of sample volume.

Protocol schematic  

Protocol schematic

Related Products

Cat. # Product Size License Quantity Details
744550.1 NucleoMag® cfDNA 48 Preps USD $373.00

The NucleoMag cfDNA kit provides an efficient, robust method for high-throughput isolation of circulating cell-free DNA (cfDNA) from 1–10 ml of human plasma obtained using EDTA or Cell-Free DNA BCT collection tubes. The kit enables recovery of fragmented DNA ≥50 bp in size via reversible adsorption of nucleic acids to paramagnetic beads under optimized buffer conditions, and eluted cfDNA is ready for downstream applications such as qPCR, ddPCR, NGS, etc. NucleoMag cfDNA is compatible with manual or automated processing and enables preparation of 24 samples in less than an hour (excluding cell lysis) on the KingFisher Flex platform.

Cat. # 744550.1 includes sufficient quantities of reagents and materials for processing 48 samples.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

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The NucleoMag cfDNA procedure

The NucleoMag cfDNA procedure

The NucleoMag cfDNA procedure. Sample lysis is achieved by incubation of plasma with Liquid Proteinase K and Lysis Buffer MCF1 at 56°C. Buffer MCF2 and paramagnetic NucleoMag P-Beads are then added, enabling optimal binding of nucleic acids. Three washing steps efficiently remove contaminating substances, such as PCR inhibitors, after magnetic separation. Residual ethanol is removed by a drying step, and highly pure DNA is eluted from the paramagnetic beads with 50–200 µl of Elution Buffer MCF5—a slightly alkaline buffer of low ionic strength (5 mM Tris/HCl, pH 8.5). The NucleoMag cfDNA kit can be used either manually or on automated liquid handling platforms or magnetic separators.

Back

Reliable purification of cfDNA from different volumes of plasma

Reliable purification of cfDNA from different volumes of plasma

Reliable purification of cfDNA from different volumes of plasma. The NucleoMag cfDNA (formerly known as NucleoMag DNA Plasma) kit was used to isolate cfDNA from 2–10 ml inputs of EDTA-treated human plasma. Analysis of purified cfDNA on the Agilent Bioanalyzer 2100 system using the High Sensitivity DNA Kit yielded a characteristic peak at 170 bp for each input volume. The total yield from each sample was determined using the PicoGreen quantification assay with the Quant-iT PicoGreen kit on a Synergy HT Multi-Detection Microplate Reader (Biotek). The consistent correlation observed between cfDNA yield and input volume demonstrates the reliable performance of the NucleoMag cfDNA kit.

Back

Efficient cfDNA recovery regardless of input volume

Efficient cfDNA recovery regardless of input volume

Efficient cfDNA recovery regardless of input volume. cfDNA isolated from varying input amounts of EDTA-treated human plasma using the NucleoMag cfDNA kit yielded clear bands upon measurement by capillary electrophoresis with the Agilent Bioanalyzer 2100 system and the High Sensitivity DNA Kit. Even for volumes as low as 2 ml, purified cfDNA was clearly visible.

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Higher, more consistent yields relative to a competitor kit

Higher, more consistent yields relative to a competitor kit

Higher, more consistent yields relative to a competitor kit. Total cfDNA was purified from 2-ml inputs of EDTA-treated human plasma from four challenging donor samples containing low levels of cfDNA (<10 ng cfDNA/ml of plasma) using NucleoMag cfDNA (formerly known as NucleoMag DNA Plasma) and a kit from Competitor T. Total DNA recovery was quantified using the Qubit dsDNA High Sensitivity kit on a Qubit fluorometer. Relative to the kit from Competitor T, NucleoMag cfDNA provided higher yields with lower sample-to-sample variability.

744550.4 NucleoMag® cfDNA 4 x 48 Preps USD $1345.00

The NucleoMag cfDNA kit provides an efficient, robust method for high-throughput isolation of circulating cell-free DNA (cfDNA) from 1–10 ml of human plasma obtained using methods such as EDTA or Cell-Free DNA BCT collection tubes. The kit enables recovery of fragmented DNA ≥50 bp in size via reversible adsorption of nucleic acids to paramagnetic beads under optimized buffer conditions. The eluted cfDNA is ready for downstream applications such as qPCR, ddPCR, NGS, etc. NucleoMag cfDNA is compatible with manual or automated processing, and enables preparation of 24 samples in less than an hour (excluding cell lysis) on the KingFisher Flex platform.

Cat. # 744550.4 includes sufficient quantities of reagents and materials for processing 192 samples.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

Documents You May Also Like Image Data

Back

The NucleoMag cfDNA procedure

The NucleoMag cfDNA procedure

The NucleoMag cfDNA procedure. Sample lysis is achieved by incubation of plasma with Liquid Proteinase K and Lysis Buffer MCF1 at 56°C. Buffer MCF2 and paramagnetic NucleoMag P-Beads are then added, enabling optimal binding of nucleic acids. Three washing steps efficiently remove contaminating substances, such as PCR inhibitors, after magnetic separation. Residual ethanol is removed by a drying step, and highly pure DNA is eluted from the paramagnetic beads with 50–200 µl of Elution Buffer MCF5—a slightly alkaline buffer of low ionic strength (5 mM Tris/HCl, pH 8.5). The NucleoMag cfDNA kit can be used either manually or on automated liquid handling platforms or magnetic separators.

Back

Reliable purification of cfDNA from different volumes of plasma

Reliable purification of cfDNA from different volumes of plasma

Reliable purification of cfDNA from different volumes of plasma. The NucleoMag cfDNA (formerly known as NucleoMag DNA Plasma) kit was used to isolate cfDNA from 2–10 ml inputs of EDTA-treated human plasma. Analysis of purified cfDNA on the Agilent Bioanalyzer 2100 system using the High Sensitivity DNA Kit yielded a characteristic peak at 170 bp for each input volume. The total yield from each sample was determined using the PicoGreen quantification assay with the Quant-iT PicoGreen kit on a Synergy HT Multi-Detection Microplate Reader (Biotek). The consistent correlation observed between cfDNA yield and input volume demonstrates the reliable performance of the NucleoMag cfDNA kit.

Back

Efficient cfDNA recovery regardless of input volume

Efficient cfDNA recovery regardless of input volume

Efficient cfDNA recovery regardless of input volume. cfDNA isolated from varying input amounts of EDTA-treated human plasma using the NucleoMag cfDNA kit yielded clear bands upon measurement by capillary electrophoresis with the Agilent Bioanalyzer 2100 system and the High Sensitivity DNA Kit. Even for volumes as low as 2 ml, purified cfDNA was clearly visible.

Back

Higher, more consistent yields relative to a competitor kit

Higher, more consistent yields relative to a competitor kit

Higher, more consistent yields relative to a competitor kit. Total cfDNA was purified from 2-ml inputs of EDTA-treated human plasma from four challenging donor samples containing low levels of cfDNA (<10 ng cfDNA/ml of plasma) using NucleoMag cfDNA (formerly known as NucleoMag DNA Plasma) and a kit from Competitor T. Total DNA recovery was quantified using the Qubit dsDNA High Sensitivity kit on a Qubit fluorometer. Relative to the kit from Competitor T, NucleoMag cfDNA provided higher yields with lower sample-to-sample variability.

744903 NucleoMag® SEP 24 Each USD $715.00

The NucleoMag SEP 24 magnetic separator is designed for the rapid manual purification of nucleic acids using the NucleoMag kits. NucleoMag kits employ Macherey-Nagel's magnetic bead-based sample processing technology, and enable rapid purification of high-quality nucleic acids from multiple samples in parallel.

The NucleoMag SEP 24 (Cat. # 744903) is used with 24-well plates.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

Documents You May Also Like Image Data

Back

744903: NucleoMag SEP 24

744903: NucleoMag SEP 24
744902 NucleoMag® SEP Maxi Each USD $264.00

The NucleoMag SEP Maxi magnetic separator is designed for the rapid manual purification of nucleic acids using the NucleoMag kits. NucleoMag kits employ Macherey-Nagel's magnetic bead-based sample processing technology, and enable rapid purification of high-quality nucleic acids from multiple samples in parallel.

The NucleoMag SEP Maxi (Cat. # 744902) is used with 50-ml tubes and carries 4 tube positions.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

Documents You May Also Like Image Data

Back

NucleoMag SEP Maxi: magnetic separator for use with 50-ml tubes

NucleoMag SEP Maxi: magnetic separator for use with 50-ml tubes

NucleoMag SEP Maxi: magnetic separator for use with 50-ml tubes.

Back

744902: NucleoMag SEP Maxi

744902: NucleoMag SEP Maxi


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