Urine Cell-Free Circulating DNA Purification Kits
Purify and concentrate high quality, high purity and inhibitor-free cell-free circulating DNA
For research use only and NOT intended for in vitro diagnostics.
Urine Cell-Free Circulating DNA Purification Kits
Purify and concentrate high quality, high purity and inhibitor-free cell-free circulating DNA
Overview
- Isolate all sizes of circulating DNA from fresh, preserved or frozen urine samples
- Isolate viral DNA
- Versatile urine input volumes
- Concentrate circulating DNA into flexible elution volumes
- Isolate inhibitor-free cell-free circulating DNA
- Compatible with Norgen’s Urine Preservative and other commercially available urine preservatives
- Purification is based on spin column chromatography that uses Norgen’s proprietary resin separation matrix
These kits provide a fast, reliable and convenient method to purify and concentrate high quality, high purity and inhibitor-free cell-free circulating DNA as well as viral DNA from fresh, preserved or frozen urine samples. All components for the purification are provided in one convenient and fast kit for the easy processing of small input volumes of bodily fluids. The purified urine DNA is fully compatible with all downstream applications including PCR, qPCR, methylation-sensitive PCR and Southern Blot analysis, Microarrays and NGS.
Background
Recent evidence indicates that cell-free circulating DNA (cf-DNA) contains valuable information for the discovery of biomarkers that can help with early detection of certain cancer types and for monitoring the disease status. The advantage for using urine as a source for cancer biomarkers is that it can be acquired in large quantities without using invasive procedures. In addition, repeated sampling from the same individual is applicable, which facilitates longitudinal studies. There are many advantages favouring the use of urinary nucleic acids for cancer biomarker discovery over blood, tissue samples or other bodily fluids, including: (1) urine is non-infectious for HIV and less infectious for many other pathogens; (2) the profile of urinary nucleic acid is similar to that in plasma or serum; (3) nucleic acid purification from urine is technically much easier because of its low protein concentration (1000-fold lower than blood). Urine cell-free circulating DNA (cf-DNA) has been utilized for the early diagnosis, prognosis and monitoring of therapy for several cancer types and autoimmune diseases, as well as for investigating fetal DNA that is normally present in the maternal blood. This cf-DNA is usually present as short fragments, generally between 150 and 250 bp or its duplicates, and is derived either from the apoptotic process, necrotic process or from the fetus.
Urine Cell-Free Circulating DNA Purification Mini Kit
For sample volumes ranging from 250 µL to 2 mL.
Urine Cell-Free Circulating DNA Purification Midi Kit
For sample volumes ranging from 2 mL to 10 mL.
Urine Cell-Free Circulating DNA Purification Maxi Kit
For sample volumes ranging from 10 mL to 30 mL.
Details
Supporting Data
Kit Specifications
|
|
Minimum Urine Input
|
10 mL
|
Maximum Urine Input
|
30 mL
|
Size of DNA Purified |
All sizes of
DNA ≥ 50 bp |
Elution Volume |
50-100 μL
|
Time to Complete 10 Purifications |
40-45 minutes
|
Average Yields |
Variable depending on specimen
|
*Please check page 6 of the product insert for 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. These kits are 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. 56600 (50 preps) | Cat. 56700 (20 preps) | Cat. 56800 (10 preps) |
---|---|---|---|
Binding Solution K | 25 mL | 75 mL | 1 x 75 mL 1 x 25 mL |
Proteinase K | 1.2 mL | 0.3 mL | 0.3 mL |
Wash Solution A | 38 mL | 2 x 38 mL | 38 mL |
Elution Buffer B | 8 mL | 15 mL | 15 mL |
Mini Spin Columns | 50 | 20 | 10 |
Midi Spin Columns | - | 20 | - |
Maxi Spin Columns | - | - | 10 |
Collection Tubes | 50 | 20 | 10 |
Elution Tubes (1.7 mL) | 50 | 20 | 10 |
Product Insert | 1 | 1 | 1 |
Documentation
FAQs
Mini, Midi, Maxi
We recommend the following steps to prepare frozen urine for isolation:
- Gently warm the sample to room temperature or 37°C for 5 min.
- DO NOT perform a centrifugation step after thawing frozen cell-free Urine samples - this will eliminate the precipitated proteins leading to loss of protein-bound cf-NA or exosomes.
- Proceed with the protocol.
We recommend the use of Norgen’s Urine Preservative when collecting urine samples. Norgen’s Urine Preservative is designed for the preservation of nucleic acids and proteins in fresh urine samples at ambient temperatures, therefore no protein precipitation will occur and the purified nucleic acids will be of a higher quality.
Citations
Title | Measurement Biases Distort Cell-Free DNA Fragmentation Profiles and Define the Sensitivity of Metagenomic Cell-Free DNA Sequencing Assays |
Journal | Clinical Chemistry. 2021. |
Authors | Chang A, Mzava O, Lenz JS, et al. |
Title | Comparison of Four Commercial Kits for Isolation of Urinary Cell-Free DNA and Sample Storage Conditions |
Journal | Diagnostics (Basel). 2020. |
Authors | Eun Young Lee, Eun-Ju Lee, Hana Yoon, Dong Hyeon Lee, and Kwang Hyun Kim |