HiPure Circulating DNA/RNA Kit

HiPure Circulating DNA/RNA Kit is designed for recovery of circulating nucleic acids from plasma, serum and other cell-free liquid samples. The kit supports column-based purification of low-abundance DNA, RNA and miRNA targets from 1–5 mL sample inputs, using a two-stage workflow that combines large-volume nucleic acid capture with micro-column concentration. This format is suitable for laboratories that need a flexible circulating nucleic acid workflow covering DNA, RNA and small RNA targets in the same product system.

The workflow is especially useful when the target is not limited to cfDNA alone. Validation testing showed recovery of low-input DNA, 50 bp short DNA fragments, miRNA-sized small RNA and RT-qPCR-compatible viral RNA from simulated plasma and water sample conditions. This makes R4316 a practical option for research workflows involving circulating DNA/RNA co-recovery, plasma miRNA analysis, viral RNA spike-in recovery or exploratory liquid biopsy sample preparation.

For laboratories focused specifically on plasma cfDNA extraction, Magen provides dedicated circulating DNA workflows with stronger product specialization. HiPure Circulating DNA Kit (IVD3182) is recommended for column-based cfDNA purification from 1–5 mL plasma or serum samples, while MagPure Circulating DNA Maxi Kit (IVD5435) provides a magnetic bead-based option for scalable cfDNA recovery and automation-oriented workflows. R4316 should therefore be selected when DNA, RNA and miRNA recovery are needed together, while IVD3182 or IVD5435 are preferred for cfDNA-focused applications.

Specifications

Principle

This kit is based on silica gel column technology. Serum or other liquid samples are lysed and digested in buffer CFL. After adding buffer CFP, the protein is removed by centrifugation to obtain the supernatant. Isopropanol is added to precipitate the total nucleic acid and transferred to the column for filtration. DNA / RNA is adsorbed on the membrane of the column, while the protein is not adsorbed and removed with the filtrate.The column is washed with buffer MGW1 to remove protein and other impurities, and then washed with buffer RW2 to remove salt. Finally, DNA / RNA is eluted by low salt buffer. The eluted DNA / RNA can be directly used for quantitative PCR/ RT-PCR, liquid or solid-phase chip analysis, hybridization and SNP detection.

Technical Validation

HiPure Circulating DNA/RNA Kit was evaluated as a column-based workflow for recovering low-abundance circulating nucleic acids from plasma and other cell-free liquid samples. The validation focused on recovery of low-input DNA, short DNA fragments, miRNA and RT-qPCR-compatible RNA targets under simulated plasma and water sample conditions.

Low-input DNA recovery was tested by adding diluted DL2000 DNA marker into 1 mL pig plasma or 1 mL RNase-free water. In plasma samples spiked with 500 ng DNA marker, R4316 recovered 333.5–337.5 ng DNA, corresponding to recovery rates of 86–87% under the tested conditions. In RNase-free water samples spiked with 20 ng DNA marker, the recovered DNA amount was 17.5–19.0 ng, corresponding to recovery rates of 86–94%. These results support efficient recovery of low-abundance DNA from both protein-containing plasma matrix and low-background water matrix.

Short-fragment recovery was evaluated using 1 mL pig plasma spiked with 50 bp DNA marker, followed by manual extraction and 50 µL elution. Nanodrop readings showed recovered nucleic acid concentrations of 48.66–58.07 ng/µL, with A260/280 values of 1.70–1.81. Agarose gel analysis showed recovery of the 50 bp DNA marker, with estimated short-fragment recovery above 80% under the tested conditions. A260/230 values were low in this experiment, which is consistent with the low nucleic acid input and guanidine-containing chemistry, so these data are used mainly as short-fragment recovery evidence rather than as a strict purity comparison.

RT-qPCR compatibility was evaluated by spiking Newcastle disease virus RNA into 1 mL pig plasma or 1 mL water sample, extracting with R4316, and analyzing the recovered RNA by fluorescent RT-PCR. R4316 extracts from plasma produced Ct values of 19.52–19.60, while water samples produced Ct values of 19.33–19.37. A reference viral nucleic acid workflow produced Ct values of 19.34 for plasma and 19.03 for water under the tested conditions. The close Ct values indicate that R4316 recovery products were compatible with RT-qPCR and did not show obvious plasma-derived inhibition in this spike-in test.

Kit Contents

Storage and Stability

The kit components can be stored at room temperature (15–25°C) and are stable for at least 18 months under these conditions.

For guidance on selecting the most appropriate DNA extraction system based on sample type, input volume and workflow requirements:

👉 Magen Kits Selection Guide

For a broader technical overview of cfDNA workflow routes, processing logic and downstream application orientation:

👉 cfDNA Extraction & Enrichment Workflows

For reference workflow structure and comparative processing logic across representative cfDNA and circulating nucleic acid workflows:

👉 cfDNA Workflow Notes

• IVD3182 — column-based cfDNA extraction workflow for routine 1–5 mL plasma or serum input.
• IVD5435 — magnetic bead-based cfDNA extraction workflow for scalable large-volume plasma or serum processing.
• 12927 — fragment enrichment workflow for short cfDNA fraction recovery and large-fragment background reduction.
• R4316 — column-based circulating DNA/RNA and RNA/miRNA workflow using large-volume capture followed by Micro Column concentration.