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 and the NEXTFLEX® Cell Free DNA Library Prep Kit for Ion Torrent™ Sequencing.

Features:

• 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

Kit Specs:

Cell Free DNA Isolation Protocol:

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