Urine Cell-Free Circulating DNA Purification Maxi Kit: Maximum Sensitivity for Large-Volume Liquid Biopsy
The Urine Cell-Free Circulating DNA Purification Maxi Kit (P4-0210) is designed for high-sensitivity applications where the detection of ultra-low abundance biomarkers is paramount. Capable of processing urine volumes from 10 mL up to 30 mL, this kit is the premier choice for researchers performing deep sequencing (NGS) or developing early-stage diagnostic assays. By increasing the starting volume, laboratories can significantly improve the probability of capturing rare circulating tumour DNA (ctDNA) or low-copy viral genomes.
The purification procedure is based on a rapid spin-column chromatography method using a proprietary resin. The process is specifically tuned to handle large volumes by utilising an initial large column to capture the nucleic acids, followed by a transition to a smaller column for final concentration and elution. This ensures the removal of all significant urinary inhibitors and contaminants, providing a purified fraction that is highly concentrated and ready for high-resolution molecular analysis. The entire workflow can be completed in approximately 45-50 minutes.
Key Benefits:
- Maximized Sample Input: Unrivalled sensitivity. Processes up to 30 mL of urine to ensure the capture of even the rarest circulating biomarkers.
- Inhibitor-Free DNA: Optimised for NGS and dPCR. Effectively removes the heavy load of inhibitors found in large-volume urine samples.
- Flexible Sample Compatibility: Works with varied sources. Suitable for fresh, frozen, or samples preserved with any major urine collection device.
- Concentrated Elution: Streamlined workflow. Elutes into 50–100 µL, eliminating the need for post-purification concentration steps.
- Reliable Performance: Highly reproducible results. Consistent yields across different donors and sample conditions.
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: P4-0210
- Manufacturer Part: 56800
- Input Volume: 10 mL to 30 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
For alternative urine input volumes, please see our other purification kit formats:
- Mini Kit (250 µL – 2 mL): View P4-0231
- Midi Kit (2 mL – 10 mL): View P4-0357