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NEXTFLEX Poly(A) Beads 2.0

The NEXTFLEX® Poly(A) Beads 2.0 kit offers a convenient method for the purification of pure, intact mRNA upstream of RNA-Seq library prep with improved mRNA yields, low rRNA contamination, and a simple protocol.

For research use only. Not for use in diagnostic procedures. 
Part Number: NOVA-512991
Format: Manual
Unit Size:
Part Number: NOVA-512992
Format: Manual
Unit Size:
Part Number: NOVA-512994
Format: Manual
Unit Size:
Part Number: NOVA-512993
Format: Manual
Unit Size:
Part Number: NOVA-512995
Format: Automated
Unit Size:
    • 10 ng – 5 µg total starting RNA
    • Magnetic bead-based protocol
    • No organic solvents or precipitation step required
    • Optimized for use with 10 ng – 5 µg of total RNA as starting material
    • Versions are available for automated and manual isolation of the mRNA transcriptome
    • Automated on the Sciclone® G3 NGSx and Zephyr® G3 NGS workstations

    Poly(A)-tailed mRNA is isolated via magnetic beads conjugated to oligo(dT), and separation using a magnetic stand allows for high mRNA recovery. The intact mRNA is eluted in small volumes, thereby eliminating the need for precipitation. These beads have been tested with the NEXTFLEX® Rapid Directional RNA-Seq Kit 2.0.

  • Reducing Ribosomal rRNA Reads in RNA-seq Libraries 

    Several approaches have been used to remove ribosomal RNA (rRNA) from total RNA samples for RNA-Seq library preparation. Removal of rRNA avoids the waste of reagents and bioinformatics resources to sequence and align ~85% of total RNA comprising rRNA (which is generally not a target of interest in NGS sequencing experiments). Revvity’s NEXTFLEX Poly(A) beads 2.0 kit offers an easy and cost-effective method for removing rRNA in RNA-Seq libraries. This product takes advantage of the tract of adenosine residues present at the 3’ ends of a vast majority of protein-coding mRNAs. Hybridization of these poly(A) tails to tracts of complementary thymidines (“oligo dT”) immobilized on solid supports allows the retrieval of poly(A)+ RNAs by recovering the solid support along with the hybridized RNA complexes.

    nextflex poly a beads 2.0 fig1

    Figure 1. Example of 1 µg of Universal Human Reference RNA (Agilent® # 740000) after Poly(A) enrichment using the NEXTFLEX Poly(A) Beads 2.0.  3 µL Poly(A) enriched RNA was run on the LabChip® GX Touch HT Nucleic Acid Analyzer using the RNA Pico Assay Reagent Kit (# CLS960012) and a DNA/RNA/Charge Variant Assay LabChip (# 760435)

    nextflex poly a beads 2.0 fig 2

    (A) mouse brain

    nextflex poly a beads 2.0 fig3

    (B) MCF7 cells

    Figure 2. NEXTFLEX Poly(A) Beads 2.0 yield clean mRNA with minimal rRNA carry-over from 5 µg of total RNA input from (A) mouse brain and (B) MCF7 cells. ~1.5% rRNA carry-over was observed based on sequencing data.

    For total RNA-Seq solution not limited to only mRNA-Seq, Revvity offers the NEXTFLEX RiboNaut rRNA depletion kit (Human / Mouse / Rat) for use NEXTFLEX Rapid Directional RNA-Seq Kit 2.0 or other applications.

  • Brand
    NEXTFLEX
    Format
    Manual
    Type
    RNA-seq