Urine Cell-Free Circulating DNA Purification Mini Kit: Rapid Recovery for Liquid Biopsies
The Urine Cell-Free Circulating DNA Purification Mini Kit (P4-0231) is an advanced solution for the rapid isolation and concentration of high-quality cell-free circulating DNA (cfc-DNA) and viral DNA from small urine volumes. Specifically designed for samples ranging from 250 µL to 2 mL, this kit addresses the challenge of recovering low-abundance biomarkers from precious or limited specimens. It provides a non-invasive alternative to traditional biopsies, facilitating the discovery of cancer biomarkers and the monitoring of disease status without the need for invasive procedures.
Utilising proprietary resin-based spin column chromatography, the kit ensures the efficient capture of all sizes of circulating DNA fragments while effectively removing urinary inhibitors such as metabolic wastes and proteins. The streamlined protocol eliminates the need for hazardous phenol-chloroform extractions or time-consuming protease treatments. Purified DNA is eluted into flexible volumes of 50 µL to 100 µL, resulting in a highly concentrated sample that is fully compatible with sensitive downstream applications, including qPCR, NGS, and methylation-sensitive PCR.
Key Benefits:
- Versatile Input Range: Optimised for small-volume samples. Effectively isolate DNA from urine volumes between 250 µL and 2 mL.
- Superior Concentration: High-affinity recovery. Concentrates cfc-DNA into small elution volumes (50–100 µL) for maximum sensitivity in molecular assays.
- Inhibitor-Free Purity: Consistent assay performance. Advanced resin technology removes PCR inhibitors, ensuring reliable results for NGS and digital PCR.
- Rapid Workflow: Accelerate your research. Process multiple samples in under 30 minutes with a simple spin-column format.
- Non-Toxic Protocol: Enhanced lab safety. No phenol/chloroform or proteases required for extraction.
Technical Insights
▼ Figure 1: cfc-DNA Purification Efficiency Across Volumes
Figure 1
Purification of cell-free circulating DNA from different urine volumes. Norgen’s Urine Cell-Free Circulating DNA Purification Mini Kit (Cat# 56600) was used to purify circulating DNA from 500 µL, 1 mL and 2 mL fresh urine. Two microlitres of the purified DNA was then used as the template in qPCR reactions to assess the relative amount of the purified housekeeping 5S rRNA gene. The relative amount of the 5S rRNA gene is linearly increasing with increasing the sample input volume. Norgen’s kit showed the most consistent and the highest recovery of the housekeeping 5S rRNA gene as compared to the other isolation method.
▼ Figure 2: Linearity of Purified DNA Recovery
Figure 2
Linearity of DNA purified from increasing urine volumes. Norgen’s Urine Cell-Free Circulating DNA Purification Mini Kit (Cat# 56600) was used to purify circulating DNA from 500 µL, 1 mL and 2 mL fresh urine. Two microlitres of the purified DNA was then used as the template in qPCR reactions to assess the linearity of the purified housekeeping 5S rRNA gene from the different urine volumes. Norgen’s Urine Cell-Free Circulating DNA Purification Mini Kit was able to recover 97% of the 5S rRNA gene from 1 mL urine relative to the amount that is present in 500 µL Urine. Moreover, 92% of the 5S rRNA gene was recovered from 2 mL urine relative to the amount that is present in 1 mL urine.
▼ Figure 3: Absence of PCR Inhibition
Figure 3
Determination of the amount of inhibition present in urine cell-free circulating DNA samples when detecting the human 5S gene. DNA was isolated from 500 µL, 1 mL and 2 mL urine using Norgen’s Urine Cell-Free Circulating DNA Purification Mini Kit (Cat# 56600). Increasing volumes of the elution (2, 4 and 8 µL) were used in a 20 µL qPCR reaction to observe any decrease in Ct value. An increase in Ct values with increasing amount of template would be a clear indication of PCR inhibitors present in the sample. An increase in elution volume used as a template in the qPCR did not affect the Ct value generated from qPCR. In fact the Ct values tend to decrease with increasing the PCR input volume, indicating that DNA purified from urine using Norgen’s kit is free of the common inhibitors usually present in urine.
Specifications
- Catalogue Number: P4-0231
- Manufacturer Part: 56600
- Input Volume: 250 µL to 2 mL
- Sample Type: Fresh, frozen, or preserved urine
- Elution Volume: 50 µL to 100 µL
- Storage Conditions: Room Temperature (15-25°C)
- Shelf Life: 2 years from date of shipment
Size Variations
Depending on your starting material volume, the Urine Cell-Free Circulating DNA Purification Kit is available in several formats:
- Midi Kit (2 mL – 10 mL): View P4-0357
- Maxi Kit (10 mL – 30 mL): View P4-0210