Urine Cell-Free Circulating DNA Purification Midi Kit: High-Performance Isolation for Medium Volumes
The Urine Cell-Free Circulating DNA Purification Midi Kit (P4-0357) is engineered for laboratories requiring high-purity isolation of cell-free circulating DNA (cfc-DNA) from urine volumes between 2 mL and 10 mL. As liquid biopsy applications expand, the ability to process larger volumes becomes critical for detecting rare genetic variants and low-concentration viral sequences. This kit provides a reliable, fast, and convenient method to concentrate high-quality, inhibitor-free DNA from fresh, preserved, or frozen urine samples.
The technology employs a sophisticated dual-column approach: an initial large-volume column handles the primary urine input, which is followed by a secondary concentration step on a mini column. This proprietary resin-based chromatography ensures that large volumes are processed efficiently while maintaining the small final elution volumes (50 µL to 100 µL) required for maximum sensitivity. The process is entirely phenol-free and does not require complex equipment, making it ideal for high-throughput biomarker discovery and clinical research.
Key Benefits:
- Extended Volume Capacity: Ideal for medium-scale inputs. Specifically optimised for 2 mL to 10 mL of urine, bridging the gap between mini and maxi formats.
- High Concentration Factor: Maximise signal detection. Effortlessly concentrate biomarkers from 10 mL of input into as little as 50 µL of elution buffer.
- Broad Molecular Weight Recovery: Captures all cfc-DNA sizes. Efficiently recovers both small apoptotic fragments and larger circulating DNA species.
- Superior Purity: Optimised for NGS. Effectively eliminates urinary salts and metabolic by-products that interfere with sequencing and PCR.
- Ambient Stability: Easy storage and handling. All components are stable at room temperature, simplifying lab logistics.
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: P4-0357
- Manufacturer Part: 56700
- Input Volume: 2 mL to 10 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
Select the kit format that best matches your experimental urine input requirements:
- Mini Kit (250 µL – 2 mL): View P4-0231
- Maxi Kit (10 mL – 30 mL): View P4-0210