Extracting cfRNA from Human Plasma or Serum

The NextPrep™ Magnazol™ cfRNA isolation kit is designed for extracting cell-free RNA (cfRNA) for constructing NGS libraries from human blood plasma or serum, using a fast magnetic bead-based format requiring minimal hands-on time. The isolated cfRNA is ideal for next generation sequencing. Suitability of the RNA for use in NGS library construction has been verified using the NEXTFLEX® small RNA-Seq v3 library prep kit for Illumina® sequencing.

Features

• Ideal for extracting cfRNA from human plasma or serum for small RNA-seq

• Magnetic beads for rapid purification of RNA from aqueous phase

• Allows highest volume of biofluids to be processed in a single tube

• Fast, < 1 hour protocol with minimal hands-on time

• Generates quality NGS libraries using few PCR cycles

• No loss of pellet during IPA precipitation

• Kit includes BCP, an alternative to chloroform

• All steps carried out at room temperature

Kit Specs

Cell Free RNA Isolation Protocol

This cell free RNA isolation procedure, which can be completed in less than 1 hour, includes an initial lysis/RNase inactivation, followed by phase separation, magnetic bead-based RNA binding step, wash steps, and elution of cfRNA from the magnetic beads.

Cell Free RNA Isolation Challenges

Extraction of cfRNA from plasma is challenging because it is highly heterogeneous in size and present in low concentrations. Since the expected yields are low, the RNA recovered cannot be reliably detected using standard methods such as NanoDrop spectrophotometer, gel or capillary electrophoresis, or RNA binding dyes. The NextPrep™ Magnazol™ cfRNA isolation kit allows for the highest volume of biofluids to be processed in a single microfuge tube compared to other kits on market.

The NextPrep™ Magnazol™ cfRNA isolation kit delivers robust cfRNA yields and quality NGS libraries with a high percentage of reads that map to miRbase than other kits on market and allows for miRNA diversity exploration using the NEXTFLEX® small RNA-Seq v3 kit.

Small RNA-seq data generated using the NEXTFLEX® small RNA-Seq v3 kit to assess miRNA mapped reads using cfRNA extracted from platelet-rich plasma using various extraction methods

KIT CONTENTS

• Magnazol™ Extraction Reagent

• Magnazol™ Magnetic Beads

• BCP (1-bromo-3-chloropropane)

• RNA Elution Solution (0.1 mM EDTA)

REQUIRED MATERIALS NOT PROVIDED

• Ethanol, 100% and 80%

• Vortex mixer

• Platform shaker, tube rotator, or heat block

• Microcentrifuge or “picofuge”

• 2 mL microfuge tubes, nuclease-free. Smaller tubes may be used for the elution step.

• Pipettors (P-1000, P-200, P-20 or equivalent) and tips

• Magnetic stand for microfuge tubes