Plasma/Serum Cell-Free Circulating DNA Purification Micro Kit: High-Yield cf-DNA Extraction from Minimal Sample Volumes
Liquid biopsies and non-invasive prenatal testing (NIPT) rely heavily on the efficient isolation of cell-free circulating DNA (cfc-DNA) from plasma or serum. However, these samples often contain extremely low concentrations of fragmented DNA and are frequently contaminated with high levels of proteins and PCR inhibitors that can compromise downstream results. The Plasma/Serum Cell-Free Circulating DNA Purification Micro Kit is specifically engineered to overcome these challenges, providing a streamlined and reliable workflow for the consistent recovery of high-quality, high-purity cfc-DNA from sample volumes as small as 10 µL.
Utilising advanced spin column chromatography, this kit employs a proprietary resin as an ion exchanger to selectively bind all sizes of circulating DNA while effectively washing away contaminating debris. This unique chemistry ensures that even the smallest DNA fragments are captured and eluted into flexible volumes ranging from 25 µL to 50 µL. The resulting purified DNA is completely free of common blood-derived inhibitors, making it an ideal template for sensitive molecular assays including NGS, qPCR, and methylation-sensitive PCR.
Key Benefits:
- Small Sample Versatility: Process 10 µL to 200 µL of input. Optimised for high-performance extraction from limited clinical or research specimens, including pediatric or rare samples.
- Comprehensive DNA Recovery: Isolate all cfc-DNA fragment sizes. Captures the full spectrum of circulating DNA, which is essential for accurate profiling in liquid biopsy and oncology research.
- Inhibitor-Free Elution: Ready for sensitive downstream assays. Superior purification chemistry removes salts, lipids, and proteins, ensuring reliable performance in NGS and qPCR without the risk of inhibition.
- Rapid and Convenient Workflow: Simple spin column procedure. Streamlines laboratory operations with a fast protocol that delivers purified DNA in less than 45 minutes without the need for phenol-chloroform extraction.
Technical Insights
▼ Technical Performance Data
Figure 1
Norgen’s Plasma/Serum Cell-Free Circulating DNA Purification Micro Kit was used to purify circulating DNA from 50 µL, 100 µL and 200 µL plasma prepared from blood collected on citrate as an anticoagulant, and compared to Competitor Q’s kit. 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 increases linearly 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
Norgen’s Plasma/Serum Cell-Free Circulating DNA Purification Micro Kit as used to purify circulating DNA from 50 µL, 100 µL and 200 µL plasma prepared from blood collected on citrate as an anticoagulant. 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 plasma volumes. Norgen’s Plasma/Serum Cell-Free Circulating DNA Purification Micro Kit was able to recover 96% of the 5S rRNA gene from 100 µL plasma relative to the amount that is present in 50 µL plasma. Moreover, 97% of the 5S rRNA gene was recovered from 200 µL plasma relative to the amount that is present in 100 µL plasma.
Figure 3
DNA was isolated from 50 µL, 100 µL and 200 µL plasma using Norgen’s Plasma/Serum Cell-Free Circulating DNA Purification Micro Kit. 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 and in fact the Ct values tend to decrease with increasing the PCR input volume indicating that DNA purified from plasma using Norgen’s kit is free of the common inhibitors usually present in plasma.
Specifications
- Catalogue Number: P4-0219
- Input Volume: 10 µL – 200 µL
- Elution Volume: 25 µL – 50 µL
- Sample Type: Fresh or frozen plasma/serum
- Prep Time: Less than 45 minutes
- Downstream Compatibility: qPCR, PCR, NGS, Southern Blot, Microarrays, Methylation-sensitive PCR