For rapid and simple purification of cell-free circulating DNA from plasma/serum samples
For research use only and NOT intended for in vitro diagnostics.
CE-IVDR marked diagnostic versions available here
For rapid and simple purification of cell-free circulating DNA from plasma/serum samples
Register today to receive an exclusive 15% off* on your first order.
These kits provide a fast, reliable and convenient spin column method for the isolation of high quality, high purity and inhibitor-free cell-free circulating DNA (cfc-DNA) from plasma/serum sample volumes. The purified plasma/serum cfc-DNA is fully compatible with all downstream applications including PCR, qPCR, methylation-sensitive PCR and Southern Blot analysis, microarrays and NGS.
Background
Plasma/Serum cell-free circulating DNA (cfc-DNA) has the potential to provide biomarkers for certain cancers and disease states as well as fetal DNA in maternal blood. Currently, significant advancements are being made in utilizing cfc-DNA as biomarkers for the early diagnosis, prognosis and monitoring of therapy for several cancer types and autoimmune diseases. Cell-free mitochondrial DNA (cfmtDNA) is also under investigation for its clinical significance. This cfc-DNA is usually present as short fragments of less than 1000 bp. In addition, cell-free fetal DNA has been widely used as a non-invasive method for prenatal diagnosis including early identification of fetal sex, genetic studies for families at high risk for inherited genetic disorders, screening for Rhesus factor, screening for aneuploidy and identification of preeclampsia.
Plasma/Serum Cell-Free Circulating DNA Purification Micro Kit
Ideal for handling initial plasma/serum volumes ranging from 10 μL up to 200 μL. The kit is designed to isolate all sizes of cfc-DNA from either fresh or frozen plasma/serum samples and the purified DNA is eluted into a flexible elution volume ranging from 25 µL to 50 µL.
Plasma/Serum Cell-Free Circulating DNA Purification Mini Kit
Ideal for handling initial plasma/serum volumes ranging from 200 μL up to 500 μL. The kit is designed to isolate all sizes of cfc-DNA from either fresh or frozen plasma/serum samples and the purified DNA is eluted into a flexible elution volume ranging from 25 µL to 50 µL.
Plasma/Serum Cell-Free Circulating DNA Purification Midi Kit
Ideal for handling initial plasma/serum volumes ranging from 1 mL - 4 mL. The first column will handle the large volume input of bodily fluids that is followed by a concentration on a mini column for a final elution of 25 µL to 50 µL
Plasma/Serum Cell-Free Circulating DNA Purification Maxi Kit
Ideal for handling initial plasma/serum volumes ranging from 5 mL- 10 mL. The first column will handle the large volume input of bodily fluids that is followed by a concentration on a mini column for a final elution of 25 µL to 50 µL
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.
Figure 4. Norgen's Plasma/Serum Cell-Free Circulating DNA Purification Mini Kit was used to purify circulating DNA from 200 µL, 300 µL and 400 µL plasma prepared from blood collected on citrate as an anticoagulant in comparison to Competitor Q's kits. 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 5. Norgen's Plasma/Serum Cell-Free Circulating DNA Purification Mini Kit was used to purify circulating DNA from 200 µL, 300 µL and 400 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 Mini Kit was able to recover 98% of the 5S rRNA gene from 300 µL plasma relative to the amount that is present in 200 µL plasma. Moreover, 98% of the 5S rRNA gene was recovered from 400 µL plasma relative to the amount that is present in 300 µL plasma.
Figure 6. DNA was isolated from 200 µL, 300 µL and 400 µL plasma using Norgen's Plasma/Serum Cell-Free Circulating DNA Purification Mini 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.
Figure 7. Purification of cell-free circulating DNA from different plasma volumes. Norgen’s Plasma/Serum Cell-Free Circulating DNA Purification Midi Kit was used to purify circulating DNA from 0.5 mL, 1 mL, 2 mL and 4 mL plasma prepared from blood collected on citrate as an anticoagulant in comparison to Competitor Q's kits. 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 8. Linearity of DNA purified from increasing plasma volumes using Norgen’s Plasma/Serum cell-free circulating DNA Purification Midi Kit. Norgen’s Plasma/Serum cell-free circulating DNA Purification Midi Kit was used to purify circulating DNA from 0.5 mL, 1 mL, 2 mL and 4 mL 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 Midi Kit was able to recover 97% of the 5S rRNA gene from 1 mL plasma relative to the amount that is present in 0.5 mL plasma. Moreover, 95% of the 5S rRNA gene was recovered from 2 mL plasma relative to the amount that is present in 1 mL plasma. Additionally, 95% of the 5S rRNA gene was recovered from 4 mL plasma relative to the amount that is present in 2 mL plasma.
Figure 9. Determination of the amount of inhibition present in plasma cell-free circulating DNA samples when detecting the human 5S gene. DNA was isolated from 0.5 mL, 1 mL, 2 mL and 4 mL plasma using Norgen’s Plasma/Serum Cell-Free Circulating DNA Purification Midi 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.
Figure 10. Norgen's Plasma/Serum cell-free circulating DNA Purification Maxi Kit was used to purify circulating DNA from 5 mL, 8 mL and 10 mL plasma prepared from blood collected on citrate as an anticoagulant in comparison to Competitor Q's kits. Two milliliters 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 11. Norgen’s Plasma/Serum Cell-Free Circulating DNA Purification Maxi Kit was used to purify circulating DNA from 5 mL, 8 mL and 10 mL plasma prepared from blood collected on citrate as an anticoagulant. Two milliliters 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 Maxi Kit was able to recover 94% of the 5S rRNA gene from 8 mL plasma relative to the amount that is present in 5 mL plasma. Moreover, 96% of the 5S rRNA gene was recovered from 10 mL plasma relative to the amount that is present in 10 mL plasma.
Figure 12. DNA was isolated from 5 mL, 8 mL and 10 mL plasma using Norgen’s Plasma/Serum Cell-Free Circulating DNA Purification Maxi 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.
| Kit Specifications | |
| Minimum Plasma/Serum Input | 10 μL |
| Maximum Plasma/Serum Input | 200 μL |
| Size of DNA Purified | ≥ 50 bp |
| Elution Volume | 25-50 μL |
| Time to Complete 10 Purifications | 15-20 minutes |
| Average Yields* | Variable depending on specimen |
*Please check page 7 in the product manual for the Plasma/Serum Average Yields and the Common DNA Quantification Methods.
Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years after the date of shipment. The kit contains a ready-to-use Proteinase K, which is dissolved in a specially prepared storage buffer. The buffered Proteinase K is stable for up to 2 years after the date of shipment when stored at room temperature.
| Component | Cat. 55500 (50 preps) | Cat. 55100 (50 preps) | Cat. 55600 (20 preps) | Cat. 55800 (10 preps) |
|---|---|---|---|---|
| Binding Buffer B | 40 mL | 2 x 40 mL | 2 x 85 mL 1 x 12 mL |
2 x 85 mL 1 x 40 mL |
| Proteinase K | 0.6 mL | 2 x 1 mL | 6.5 mL | 8 mL |
| Solution WN | 18 mL | 18 mL | 18 mL | 18 mL |
| Wash Solution A | 18 mL | 38 mL | 18 mL | 18 mL |
| Elution Buffer B | 8 mL | 8 mL | 30 mL | 30 mL |
| Micro Spin Columns | 50 | 50 | 20 | 10 |
| Mini Spin Columns | - | 50 | - | - |
| Midi Spin Columns | - | - | 20 | - |
| Maxi Spin Columns | - | - | - | 10 |
| Collection Tubes | 50 | 100 | 20 | 10 |
| Elution Tubes (1.7 mL) | 50 | 100 | 20 | 10 |
| Product Insert | 1 | 1 | 1 | 1 |
| Title | Exploring mitochondrial DNA copy number in circulating cell-free DNA and extracellular vesicles across cardiovascular health status: A prospective case-control pilot study |
| Citation | The FASEB journal 2024. |
| Authors | Chiara Rucci1,2 | Gaia de Simone1,2 | Saniya Salathia3 | Cristina Casadidio3 | Roberta Censi3 | Laura Bordoni |
| Title | Monitoring of plasma circulating donor DNA reflects cardiac graft injury: Report of two cases |
| Citation | BIOMEDICAL REPORTS 2024. |
| Authors | DANA DLOUHA1 , PAVLINA HUCKOVA1 , EVA ROHLOVA1-3, JEVGENIJA VYMETALOVA4 , SARKA NOVAKOVA1 and JAROSLAV A. HUBACEK |
| Title | Plasma cfDNA abundance as a prognostic biomarker for higher risk of death in geriatric cardiovascular patients |
| Citation | Mechanisms of Ageing and Development 2024. |
| Authors | Maurizio Cardelli a, Francesca Marchegiani b, Pierpaolo Stripoli b, Francesco Piacenza a, Rina Recchioni b, Mirko Di Rosa c, Robertina Giacconi a, Marco Malavolta a, Roberta Galeazzi b, Beatrice Arosio d e, Fiammetta Cafarelli f, Francesco Spannella g h, Antonio Cherubini i, Fabrizia Lattanzio j, Fabiola Olivieri |
| Title | Urine-Based Biomarker Test Uromonitor® in the Detection and Disease Monitoring of Non-Muscle-Invasive Bladder Cancer— A Systematic Review and Meta-Analysis of Diagnostic Test Performance |
| Citation | Cancers 2024. |
| Authors | Anton P. Kravchuk 1,†, Ingmar Wolff 2,*,†, Christian Gilfrich 1, Ralph M. Wirtz 3, Paula Soares 4,5, Kay-Patrick Braun 6, Sabine D. Brookman-May 7,8, Lisa Kollitsch 9, Katharina Hauner 10, Martin Burchardt 2, Johannes Bründl 11, Maximilian Burger 11,‡ and Matthias May 1,‡ |
| Title | Abstract 6012: Comparison of pre-analytical methods for DNA methylation analysis of cfDNA |
| Citation | Cancer Res 2023. |
| Authors | Miljana Tanic; Oluwadunni Emiloju; Pawan Dhami; Stephan Beck |
| Title | Are elevated mitochondrial DNA fragments in prostatic inflammation a potential biomarker for prostate cancer? |
| Citation | Original Papers - Prostate 2023. |
| Authors | Ugur Aferin, Nurten Bahtiyar, Ilhan Onran & Hamdi Ozkara |
| Title | Multidimensional fragmentomic profiling of cell-free DNA released from patient-derived organoids |
| Citation | Human genomics 2023. |
| Authors | Jaeryuk Kim, Seung-Pyo Hong, Seyoon Lee, Woochan Lee, Dakyung Lee, Rokhyun Kim, Young Jun Park, Sungji Moon, Kyunghyuk Park, Bukyoung Cha & Jong-Il Kim |
| Title | Significance of Circulating Cell-Free DNA Biomarkers in HBeAg-Negative Chronic Hepatitis B Virus Infection and Their Changes after Treatment Initiation |
| Citation | Pathogens 2023. |
| Authors | Nikolaos D. Karakousis ,Lampros Chrysavgis,Alkistis Papatheodoridi, Aigli-Ioanna Legaki ,Panagiotis Lembessis ,Evangelos Cholongitas ,Antonios Chatzigeorgiou, and George Papatheodoridis |
| Title | Application and optimization of minimally invasive cell-free DNA techniques in oncogenomics |
| Citation | Tumor Biology 2018. |
| Authors | Kumar, M., Choudhury, Y., Ghosh, S. K., & Mondal, R. (2018). |
| Title | Immunohistochemical evaluation of stressresponsive protein sestrin2 and its correlation with p53 mutational status in eyelid sebaceous gland carcinoma |
| Citation | British Journal of Ophthalmology 2018. |
| Authors | Jayaraj, P., Sen, S., Rangarajan, S., Ray, N., Vasu, K., Singh, V. K., ... & Verma, A. (2018). |
