Figure 1. The Apostle MiniMax™ kit was used to extract DNA from 1 mL of plasma collected in 3 types of stabilization containers, referred to here as cfDNA Tube 1, EDTA Collection Tube, and cfDNA Tube 2. A market leading column-based kit (Kit A) was also used to extract DNA from 1 mL of plasma from cfDNA Tube 1, EDTA Collection Tube, and cfDNA Tube 2. A second bead-based kit (Kit B) was used to extract DNA from the EDTA Collection Tube. DNA yield was quantified using Quant-iT PicoGreen dsDNA assay kit (Thermo Fisher Scientific). The error bars represent the standard deviation of three technical replicates. For each tube type, the Apostle MiniMax™ kit extracted a higher total yield of DNA. DNA size was analyzed on an Agilent Bioanalyzer 2100. For all three tube types the Bioanalyzer traces indicate the DNA extracted correlates with the expected sizes of cfDNA (~170 bp).

Figure 2. Left: DNA isolation from varying plasma amounts. Nuceleosomes were spiked into plasma to ensure enough DNA was present for QC. The Apostle MiniMax™ kit isolated nearly all the input DNA, while Kit 2 isolated less from larger plasma volumes. Right: Isolation of DNA from 1–5 mL of EDTA plasma. Yields were highest with the Apostle MiniMax™ kit for 3-5 mL, and yields were similar across all kits at 1 mL.

Figure 3. DNA was extracted from 1 mL of plasma collected in cfDNA Tube 1 using the Apostle MiniMax™ kit and one other kit (Kit A). These are technical replicates and both replicates extracted using the Apostle MiniMax™ kit are nearly identical. In contrast, the replicates extracted using Kit A have two distinct traces. Isolated cfDNA was characterized on an Agilent Bioanalyzer 2100; the peaks at 35 and 10380 bp are internal markers.

Figure 4. Left: 20 μL of a 170 bp synthetic DNA fragment containing the EGFR C2573T>G L858R mutation was spiked into 1 mL of TE buffer (blue) or serum (red) at concentrations ranging from 1 ng/μL to 0.001 ng/μL. The Apostle MiniMax™ kit was used to isolate the DNA fragments. A) The qPCR amplification plot shows highly overlapping curves for the isolated DNA fragments and the original synthetic DNA solution at the different concentrations. B) To quantify the amount of DNA that Apostle MiniMax™ kit can recover, a qPCR standard curve was generated using the original synthetic DNA solution. The DNA isolation recovery rate was calculated to be >90% for the Apostle MiniMax™ kit. Right: qPCR performance was compared between Apostle MiniMax™ kit and one other market leading column-based kit (Kit A). Apostle MiniMax™ kit outperformed Kit A for DNA extracted from all three collection tubes presented.