Urine Cell-Free Circulating DNA Purification Midi Kit

  • Isolate all sizes of circulating DNA from fresh, preserved or frozen urine samples
  • Isolate viral DNA
  • Versatile urine input volumes (2 mL - 10 mL)
  • Concentrate circulating DNA into a flexible elution volume ranging from (50 µL - 100 µL)
  • Isolate inhibitor-free cell-free circulating DNA
  • Purify high-quality DNA in 40-45 minutes
  • Compatible with Norgen’s Urine Preservative and other commercially available urine preservatives
Items Cat. # Size
Urine Cell-Free Circulating DNA Purification Midi Kit 56700 20 preps

Urine Cell-Free Circulating DNA Purification Midi Kit

This kit provides 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 from volumes ranging from 2 mL to 10 mL. The first column will handle the large volume input of urine that is followed by a concentration on a mini column for a final elution of 50 µL to 100 µL. All components for the purification are provided in one convenient & fast kit for the easy processing of small input volumes of bodily fluids. For a schematic workflow of the protocol click here.

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.

Protocol

Workflow

Kit Specifications
Minimum Urine Input
2 mL
Maximum Urine Input
10 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 buffers should be kept tightly sealed and stored at room temperature (15-25°C) for up to 2 years without showing any reduction in performance.

Norgen’s Urine Cell-Free Circulating DNA Purification Kits contain ready-to-use Proteinase K solution, which is dissolved in a specially prepared storage buffer. The Proteinase K is stable for up to 2 years after delivery when stored at room temperature. To prolong the lifetime of Proteinase K, storage at 2–8°C is recommended.

 

 

Kit Components
Component Cat. 56700 (20 preps)
Binding Solution K 75 mL
Lysis Buffer A 20 mL
Proteinase K 0.3 mL
Wash Solution A 2 x 38 mL
Elution Buffer B 15 mL
Mini Spin Columns 20
Midi Spin Columns 20
Collection Tubes 20
Elution Tubes (1.7 mL) 20
Product Insert 1