From lysis through to separation, the protocol can be fully automated and is compatible with most open-source liquid handlers.

High binding capacity beads maximise cfDNA recovery and an elution volume as low as 30 µl ensures cfDNA is concentrated enough for downstream applications without the need for DNA vacuum concentration.

The kit uses the reversible nucleic acid-binding properties of paramagnetic particles along with an optimised binding system that targets recovery of small cfDNA fragments and minimises gDNA contamination.

Bead Xtract cfDNA, 20 (1 ml) extractions

 Bead Xtract cfDNA, 200 (1 ml) extractions

Bead Xtract cfDNA, 800 (1 ml) extractions

The high binding capacity of the paramagnetic particles used in the Nonacus Bead Xtract cfDNA kit maximises cfDNA yield from plasma samples. It also reduces the volume of particles required to isolate the cfDNA so that an elution volume as low as 30 ul can be achieved even with very large volumes of plasma.

We compared the DNA extraction yield (ng) of the Bead Xtract cfDNA kit to a range of spin-column and bead-based cfDNA extraction kits currently available on the market.  DNA extraction was performed manually using 1 ml plasma replicates following the individual company instructions and DNA quantification showed that the Bead Xtract cfDNA kit outperformed the other extraction methods (Figure 1).

The Bead Xtract cfDNA kit and protocol are optimized for recovering smaller DNA fragments ensuring that cfDNA yield is maximized. Figure 2 shows typical nucleosomal patterning indicative of successful cfDNA isolation and minimal gDNA contamination. This ensures that high quality cfDNA is recovered from your samples enabling more sensitive detection of genomic variants in downstream applications like qPCR, ddPCR and NGS.

Figure 1. Average DNA yield (ng) extracted from 1 ml plasma replicates using a range of different bead-based and spin-column based extraction kits. cfDNA was extracted following the manufacturer's instructions and eluted into 50 ul of elution buffer. DNA concentration was determined using high sensitivity reagents on the Qubit 3.0 (Invitrogen).

Figure 2. Automated cfDNA extraction using the Bead Xtract cfDNA kit shows expected nucleosomal patterning typical of cfDNA fragment distribution and low gDNA contamination.    4 ml replicates performed on the Hamilton ML STAR and eluted in 50 µl of buffer. Purified DNA was analysed on the Agilent 4200 Tapestation.

The properties of the paramagnetic particles in the Bead Xtract cfDNA kit ensure fast magnetic separation even when using large volumes of plasma.

The elimination of funnels and vacuum steps streamlines the process so that manual extraction takes less than 70 minutes with a hands-on time of under 30 minutes.

The Bead Xtract cfDNA protocol offers users flexibility to be run manually or as a fully automated process to suit laboratory and sample throughputs. Validation of automated isolation from lysis to extraction has been performed on the Hamilton ML STAR but protocols and scripts are available for a range of open-ended liquid handlers. Please contact //nonacus.com/bead-xtract-cfdna/support@nonacus.com" style="box-sizing: border-box; background-color: transparent; color: rgb(0, 0, 0); line-height: 1.5;">support@nonacus.com for more information.

Fully automated cfDNA extraction requires just 30 minutes set-up time and has been shown to generate highly concordant results with manual processing.