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Flexible magnetic bead based purification of cell-free DNA from 1–10 mL blood plasma
| Application | Isolation of cfDNA |
| Selling unit | 48 Preps, 192 Preps |
| Target | DNA |
| CE certified | No, research use only |
| Technology | Magnetic bead technology |
| Brand | NucleoMag |
| Format | Magnetic beads |
| Handling | Magnetic separation |
| Automated use | Yes |
| Automation support | Compatible with most open robotic platforms. Please visit www.mn-net.com/automation or consult our technical support for further information. |
| Sample material | Cell-free BCT (Streck), Plasma, Serum |
| Sample amount | 1−10 mL |
| Fragment size | > 50 bp |
| Theoretical binding capacity | 0.3 µg/µL beads |
| Typical purity A260/A280 | 1.6–1.9 |
| Elution volume | 50–200 µL |
| Preparation time | 60 min/24 preps (excl. lysis) |
| Typical downstream application | ddPCR, NGS, qPCR |
| Storage temperature | 15–25 °C / 59–77 °F |
| Hazardous material | Yes |
The NucleoMag cfDNA kit is designed for the rapid manual and automated isolation of circulating cell-free DNA from human plasma, derived from e.g., EDTA blood draw tubes or Cell-Free DNA BCT. The kit can be processed with the NucleoMag SEP 24.
The isolation of cfDNA from human EDTA plasma using the NucleoMag cfDNA kit shows clear bands after measurement by capillary gel electrophoresis using the Agilent Bioanalyzer 2100 system with the High Sensitivity DNA kit. Even for low volumes such as 2 mL the detection of cfDNA is clearly visible.
| Plasma volume | Total cfDNA recovery (mean value) |
|---|---|
| 2mL | 12.0ng |
| 4mL | 17.5ng |
| 6mL | 28.5ng |
| 8mL | 39.0ng |
| 10mL | 50.5ng |
Total cfDNA from 2 mL human EDTA plasma derived from 4 challenging donor samples with low abundance cfDNA (< 10 ng cfDNA/ mL Plasma) was purified. Isolation with the NucleoMag cfDNA kit results in higher and more consistent total cfDNA yields with less fluctuations in comparison to the competitor T. The final total DNA recovery was quantified using the Qubit dsDNA High Sensitivity kit (ThermoFisher Scientific) on a Qubit fluorometer (ThermoFisher Scientific).
The NucleoMag cfDNA kit was used to isolate cfDNA from human EDTA plasma samples, ranging from 2–10 mL. Efficient purification of cfDNA is shown by the characteristic peak at approx. 170 bp using the Bioanalyzer 2100 system and the High Sensitivity DNA Kit from Agilent. The total yield was determined by PicoGreen quantification assay using the Quant-iT PicoGreen kit (ThermoFisher Scientific) on a Synergy HT Multi-detection microplate reader (Biotek). The consistent correlation of cfDNA amount and sample volume demonstrates reliable performance independent on sample volume.
