FOR ISOLATION OF CIRCULATING DNA FROM URINE FOR NGS
NextPrep-Mag™ Urine cfDNA Isolation Kit
- Ideal for extracting DNA from urine for cell-free DNA analysis
- Generates highly concentrated libraries using few PCR cycles
- Automation friendly, mag-bead based format
- Fast, 30 minute protocol with minimal hands-on time
- Ideal for NGS library prep
- No vacuum pumps, vacuum manifolds, or column extenders required
Extracting cfDNA from Urine
The NextPrep-Mag™ Urine cfDNA Isolation Kit is designed for extracting cell-free DNA (cfDNA) for constructing NGS libraries from urine, using a fast magnetic bead-based format requiring minimal hands-on time. The isolated cell free DNA is ideal for next generation sequencing and PCR analysis. Suitability of the DNA for use in NGS library construction has been verified using the NEXTFLEX® Cell Free DNA-Seq Library Prep Kit for Illumina® Sequencing Platforms.
NextPrep-Mag™ Urine cfDNA Isolation Kit Compared to the QIAamp™ Circulating Nucleic Acid Kit
The NextPrep-Mag™ Urine cfDNA Isolation Kit offers a number of advantages over the QIAamp™ Circulating Nucleic Acid Kit, including speed of cfDNA isolation and ease of automation, while delivering comparable cfDNA yields and higher concentration NGS libraries.
Cell Free DNA Isolation Protocol
The procedure, which can be completed in approximately 30 minutes, includes an initial lysis and DNA binding step, wash steps, and elution of cfDNA from the magnetic beads. A distinguishing feature of the kit is the very rapid magnetic bead attraction steps.
URINE + BINDING SOLUTION + PROTEINASE K + MAGNETIC BEADS (15 MIN)
ATTRACT TO MAGNET & REMOVE FLUID
RAPID WASH STEPS
ELUTE (5 MIN)
TRANSFER cfDNA
KIT CONTENTS
- cfDNA DNA Proteinase K
- Urine cfDNA Binding Solution
- cfDNA Magnetic beads
- cfDNA Wash Solution 1
- cfDNA Wash Solution 2
- cfDNA Elution Solution
- Control PCR Primers
REQUIRED MATERIALS NOT PROVIDED
- Ethanol, 100%
- Vortex mixer
- Heat block at 55°C
- Magnetic stands to hold 2 mL microcentrifuge tubes and 15 mL or 50 mL tubes
- Microcentrifuge or low-speed “picofuge”
- Nuclease-free 2 mL (1.5 mL and 0.5 mL optional) microcentrifuge tubes and 15 mL tubes
- Pipettors (P-1000, P-200, P-20) and tips
- Disposable pipettes (variable sizes depending on sample volume processed)
Selected Citations that Reference the Use of the NextPrep-Mag Urine cfDNA Isolation kit:
Streleckiene, G., Reid, H. M., Arnold, N., Bauerschlag, D., & Forster, M. (2018). Quantifying cell free DNA in urine: Comparison between commercial kits, impact of gender and inter-individual variation. BioTechniques,64(5), 225-230. doi:10.2144/btn-2018-0003
The NextPrep-Mag™ Urine cfDNA Isolation Kit For < 1 mL – 4 mL contains sufficient reagents for extraction of DNA from 50 samples of 1 mL urine or less, or 12 samples of 4 mL of urine. The NextPrep-Mag™ Urine cfDNA Isolation Kit For 4 mL – > 20 mL contains sufficient reagents for extraction of DNA from 25 samples of 4 mL urine or less, or 5 samples of 20 mL of urine. The kit components are stable for at least 12 months from time of receipt, when properly stored.