Apostle MiniMax™ Performance and Data
Cell-Free DNA Isolation from Plasma for Liquid Biopsy
Apostle MiniMax™ High Efficiency cfDNA Isolation Kit (Standard Edition) is a cell-free DNA isolation reagent kit built on magnetic bead-based technology. Apostle MiniMax™ has been demonstrated to purify cfDNA from human plasma in both manual and automated workflows.
- Data representative of results from 1-5 ml of plasma
- Demonstrated compatibility with a variety collection tubes
- cfDNA purity shown to be suitable for downstream PCR based assays
Plasma Collected in a Variety of Collection Tubes
Figure 1 (below): DNA was extracted from 1 mL of plasma collected in cfDNA Tube 1 and cfDNA Tube 2 using Apostle MiniMax one other market leading column based kit (kit A). DNA was also extracted from 1 mL of plasma collect in EDTA collection tubes using Apostle MiniMax one other market leading column based kit (kit A) and a second bead based kit (kit B). 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 the Apostle MiniMax extracted higher total yield of DNA. DNA size was analyzed on an Agilent Bioanalyzer 2100. For all three tube types the Bioanlyzer traces indicate the DNA extracted correlates with the expected sizes of cfDNA (~170 bp).
Figure 2 (below): DNA was extracted from 1 mL of plasma collected in in cfDNA Tube 1 using Apostle MiniMax™ and one other kit. These are technical replicates and both of the replicates that were extracted using Apostle MiniMax™ are exactly the same, while the DNA extracted using Kit A have two distinct traces. Isolated cfDNA was characterized on Agilent Bioanalyzer 2100; the peaks at 35 and at 10380 correspond to internal markers.
In Figure 3 (data set A) cfDNA was isolated from 4 mL of cell-free plasma with Apostle MiniMax (red curve) and market leading column based kit (blue curve). The isolated cfDNA was characterized by Bioanalyzer 2100. In data set B cfDNA was isolated from 20 mL of cell-free urine using the Apostle MiniMax (red curve) and market leading column based kit (blue curve). The isolated cfDNA was characterized by Bioanalyzer 2100 as indicated by higher peaks at ~170bp. The peaks at 35 and 10380 correspond to internal markers. For both plasma and urine samples Apostle MiniMax outperformed the major alternative product.
DNA Mutation Detection and qPCR Performance
In Figure 4 (below 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. Apostle MiniMax™ was used to isolate the fragment: A) is the amplification plot, which shows highly overlapping curves for the isolated mutated DNA fragment and the original DNA solution at the different concentrations, and B) quantify the amount of DNA that Apostle MiniMax™ can recover a qPCR standard curve was generated using the original DNA. DNA isolation recovery rate was calculated to be >90%. In Figure 5 (below right) qPCR performance was compared between Apostle MiniMax™ and one other market leading column based kit (Kit A). Apostle MiniMax™ outperformed Kit A for DNA extracted from all three collection tubes presented.
DNA Yield Increases with Input Volume
In Figure 6 (below) DNA was extracted from 1, 3, and 5 mL of plasma collected in EDTA tubes Apostle MiniMax™. DNA yield increases with the amount of input if using Apostle MiniMax™. DNA yield was quantified by Quant–iT PicoGreen dsDNA assay kit (Thermo Fisher Scientific). Error bars represent the standard deviation of three technical replicates.
- Lyse plasma
- Bind cfDNA to magnetic beads
- Separate magnetic beads from contaminants
- Wash magnetic beads with Wash Solution
- Wash magnetic beads with 2nd Wash solution
- Elute cfDNA from magnetic beads
Products: Apostle MiniMax™ High Efficiency cfDNA Isolation Kit (Standard Edition)
|C40605||Apostle MiniMax™ Large Volume||50|
Evaluate Apostle MiniMax™
Interested in a sample kit? Evaluate Apostle MiniMax™ High Efficiency Cell-Free DNA Isolation Kit today. The kit isolates cell-free DNA (cfDNA) from plasma, serum, and urine, and is powered by magnetic bead-based technology suitable for NGS and PCR-based downstream applications.
Beckman Coulter makes no warranties of any kind whatsoever express or implied, with respect to this protocol, including but not limited to warranties of fitness for a particular purpose or merchantability or that the protocol is non-infringing. All warranties are expressly disclaimed. Your use of the method is solely at your own risk, without recourse to Beckman Coulter. Not intended or validated for use in the diagnosis of disease or other conditions. This protocol is for demonstration only, and is not validated by Beckman Coulter.
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