Takara NucleoMag cfDNA \4 x 48 Preps

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.

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.

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.

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.

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.