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  • NEXTFLEX Rapid XP V2 DNA-Seq Kit

NEXTFLEX Rapid XP V2 DNA-Seq Kit

NEXTFLEX NGS Library Prep
NEXTFLEX Rapid XP V2 DNA-Seq Kit
NEXTFLEX NGS Library Prep
Revvity kit box for reagents and consumables
NEXTFLEX NGS Library Prep

The NEXTFLEX™ Rapid XP V2 DNA-Seq Kit is a fragmentation-based whole genome sequencing (WGS) library preparation workflow, taking samples from DNA to sequencing-ready libraries in as little as 2.5 hours. Proprietary NEXTFLEX normalization beads equalize library concentrations during the library prep process, so no separate quantitation or manual pooling is required.

The protocol supports inputs from 100 pg to 1 µg and offers a PCR-free option for ≥ 250 ng, preserving library complexity and minimizing GC bias. Resulting libraries show uniform coverage and low duplicate rates on Illumina® and Element Biosciences™ AVITI platforms, enabling confident variant detection in whole genome, metagenomic, and other research applications.

Automation-ready reagents, high indexing capacity (up to 1,536 UDIs), and streamlined clean-ups deliver fast, reproducible DNA-seq results at any scale.

View product information ビュー related reagents
Feature Specification
Automation Compatible Yes
Product Group DNA-seq

The NEXTFLEX™ Rapid XP V2 DNA-Seq Kit is a fragmentation-based whole genome sequencing (WGS) library preparation workflow, taking samples from DNA to sequencing-ready libraries in as little as 2.5 hours. Proprietary NEXTFLEX normalization beads equalize library concentrations during the library prep process, so no separate quantitation or manual pooling is required.

The protocol supports inputs from 100 pg to 1 µg and offers a PCR-free option for ≥ 250 ng, preserving library complexity and minimizing GC bias. Resulting libraries show uniform coverage and low duplicate rates on Illumina® and Element Biosciences™ AVITI platforms, enabling confident variant detection in whole genome, metagenomic, and other research applications.

Automation-ready reagents, high indexing capacity (up to 1,536 UDIs), and streamlined clean-ups deliver fast, reproducible DNA-seq results at any scale.

View product information ビュー related reagents
Product variants
Unit Size: 8 rxns
Part #:
NOVA-5149-21
Unit Size: 48 rxns
Part #:
NOVA-5149-22
Unit Size: 96 rxns
Part #:
NOVA-5149-23
For research use only. Not for use in diagnostic procedures.
Request more information
NEXTFLEX NGS Library Prep
NEXTFLEX Rapid XP V2 DNA-Seq Kit
NEXTFLEX NGS Library Prep
Revvity kit box for reagents and consumables
NEXTFLEX Rapid XP V2 DNA-Seq Kit
NEXTFLEX NGS Library Prep

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Product information

  • Overview
  • Additional product information
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Overview

  • Single-tube enzymatic fragmentation, end-repair & A-tailing - DNA to sequencing-ready libraries in as little as 2.5 hours
  • PCR-free option for ≥ 250 ng input plus a broad 100 pg - 1 µg range for low- and high-input samples with PCR amplification
  • Built-in NEXTFLEX normalization beads - skip post-library quantitation and manual pooling
  • Ultra-low adapter-dimer formation & uniform GC coverage for confident variant detection
  • Up to 1,536 UDIs available for large, multiplexed studies
  • 96 UDI-UMI barcode set enables molecular error-correction for rare variant calling
  • Automation-ready workflow validated on Revvity Automation NGS workstations
  • Quality assured through rigorous testing of every lot

The NEXTFLEX Rapid XP V2 DNA-Seq Kit unites speed, data quality, and scalability: enzymatic fragmentation streamlines the workflow, proprietary normalization beads remove an entire QC step, and an optional PCR-free path preserves library complexity. Whether processing a handful of samples or a 96-well plate on a liquid-handler, you get consistent, unbiased libraries ready for Illumina® and Element AVITI™ sequencing.

Additional product information

One-tube enzymatic fragmentation - libraries in as little as 2.5 hours

The NEXTFLEX enzyme cocktail performs fragmentation, end-repair, and A-tailing in a single tube. A complete manual run, from DNA to normalized libraries, takes as little as 2.5 hours total. Fragment size is tunable by adjusting the 35°C incubation. This streamlined chemistry removes the need for sonication equipment, cuts out an extra cleanup step, and reduces thermocycler runs.

Broad input range WGS with PCR-free option

The protocol accepts 100 pg – 1 µg of genomic DNA. For ≥ 250 ng inputs, users may skip PCR (Appendix A workflow), preserving library complexity and minimizing duplicates; low-input samples use 2–15 cycles as recommended. Expected post-PCR yield is 100–500 ng, sufficient for one or more flow-cell lanes.

Highly Efficient Ligation with Minimal Adapter-Dimer Formation

The NEXTFLEX Rapid XP V2 DNA-Seq Kit delivers exceptional ligation efficiency in a single 20°C incubation.

  • 30-minute incubation – keeps total library-prep time at 2.5 hours for time-sensitive runs.
  • 60-minute incubation – produces optimal ligation efficiency when the highest possible conversion is critical, for example, PCR-free workflows or ultra-low-input samples.

Optimized reagents join adapters without a prior dilution step, converting nearly all fragments across our entire input range. Electropherograms from 500 pg and 1 ng libraries (Figure 1) show no detectable adapter-dimer peak (~150 bp), even at picogram inputs. This combination of high ligation yield and low dimer formation increases usable sequencing reads and ensures reliable, reproducible data.

Electropherogram of 500 pg and 1 ng libraries prepared with the NEXTFLEX Rapid XP V2 DNA-Seq Kit: single-tube enzymatic fragmentation and 60-min ligation yield clean ~425 bp peaks with no detectable ~150 bp adapter-dimer signal.

Figure 1. Electropherogram traces for libraries prepared from 500 pg (A) and 1 ng (B) human gDNA with the NEXTFLEX Rapid XP V2 DNA-Seq Kit. The dominant library peak is flanked by an adapter-dimer region at ~150 bp that is virtually absent, confirming highly efficient ligation and cleanup even at sub-nanogram inputs.

Uniform coverage across the GC spectrum

GC-bias analysis shows flat, normalized depth from ~20% to 60% GC, independent of input mass; only mild drop-off appears above 60%. This consistency yields near-complete genome coverage at standard read depths, boosting SNP/indel sensitivity without expensive over-sequencing.

Line graph showing normalized read depth versus GC content using NEXTFLEX Rapid XP V2 DNA-Seq; curves stay relatively uniform from 20–60 % GC indicating minimal bias

Figure 2. Normalized coverage across varying %GC content for the 8 replicates analyzed. The minimal GC bias observed indicates a high degree of coverage uniformity in the data generated.

Built-in NEXTFLEX Normalization Beads - save ≈ 3 h per 96-sample batch

Proprietary beads bind a fixed mass of library DNA during the final cleanup, so every sample leaves the protocol at the same molarity. This single step simplifies the workflow, reduces hands-on time, and ensures optimal library-input balance for precise pooling and sequencing. By eliminating post-library qPCR/fluorometric quantitation and manual pooling, labs can save up to three hours on a 96-sample run. The on-bead normalization also cuts sample-to-sample variability, preventing costly resequencing due to over- or under-clustering and eliminates the need for quantitation reagents and instrumentation.

Flowchart comparing traditional library normalization (quantify, size, dilute & pool) with NEXTFLEX Rapid XP V2 on-bead normalization that adds no extra workflow time.

Figure 3. Workflow comparison of traditional post-library normalization versus built-in NEXTFLEX bead normalization. The conventional route requires three downstream QC steps, quantify libraries, fragment sizing, dilute & pooling, adding ~3 hours to turnaround, while NEXTFLEX beads perform normalization during library prep with no extra time.

Indexing options - from duplex-error correction to population-scale multiplexing

NEXTFLEX indexing chemistry is modular, letting you choose the depth of error control and the level of multiplexing your project demands:

Option Core benefit Ideal use-cases
UDI-UMI plates (10 bp UDI + 9 bp UMI) Single-molecule quantification & bias-free duplicate removal cfDNA/liquid-biopsy panels, ultra-deep exome, antimicrobial-resistance screens
High-plex UDI sets (8–10 bp UDI) Massive multiplexing with zero index reuse, perfect for patterned-flow-cell sequencers. Population-scale WGS, large metagenomic surveys, core-facility batching

 

Automation-ready workflows

Revvity provides vendor-qualified scripts for Sciclone™ G3 NGSx/iQ, Zephyr™ NGS, and Fontus™ workstations, so users can drop the NEXTFLEX onto pre-mapped deck positions and run a validated protocol - no custom coding or tip-mapping required.

Specifications

Automation Compatible
Yes
Product Group
DNA-seq
Shipping Conditions
Dual Temperature
Unit Size
8 rxns

Citations

  • Gaulke, C. A., Schmeltzer, E. R., Dasenko, M., et al. (2021). Evaluation of library preparation procedure and sample characteristics on metagenomic profiles. mSystems, 6(5), e00440-21.
  • Author(s). (2024). Beyond single diagnosis: Exploring multidiagnostic realities in [Journal Name]. https://doi.org/10.1155/2024/9115364​
  • Di Martino M L, Jenniches L, Bhetwal A, Eriksson J, Lopes A C C, Ntokaki A, Pasqua M, Sundbom M, Skogar M, Graf W, Webb D-L, Hellström P M, Mateus A, Barquist L, & Sellin M E. (2025). A scalable gut epithelial organoid model reveals the genome-wide colonization landscape of a human-adapted pathogen. Nature Genetics. Advance online publication. https://www.nature.com/articles/s41588-025-02218-x

FAQs

  • How long does library prep take with the NEXTFLEX Rapid XP DNA-seq kit v2?

    A manual workflow (benchtop incubations + bead clean-ups) takes as little as 2.5 hours from DNA to normalized libraries.

  • Do I still need mechanical shearing or tagmentation?

    No. The kit includes a single-tube enzymatic reaction that fragments, end-repairs, and A-tails DNA in one step, eliminating sonication.

  • What DNA input range is supported, and when can I skip PCR?

    The protocol works from 100 pg to 1 µg. Inputs of ≥ 250ng can omit PCR entirely (PCR-free appendix), while lower inputs require 2–15 PCR cycles.

  • How do Normalization Beads save time?

    The beads bind a fixed mass of library DNA during the final cleanup, so labs skip qPCR/fluorometric quantitation and manual pooling — saving up to three hours per 96-sample run.

  • How uniform is coverage across different GC contents?

    Normalized read-depth curves stay within relatively uniform from 20% to 60% GC across inputs from 1 ng to 750 ng, indicating minimal GC bias.

  • What level of adapter-dimer formation should I expect?

    Electropherograms of 500 pg and 1 ng libraries show no detectable adapter-dimer peak (~150 bp).

  • Which sequencing platforms are compatible?

    Libraries are ready for all Illumina® instruments and Element Biosciences AVITI™ systems.

  • Do I still use Normalization Beads if follow PCR-free Protocol? 

    Normalization beads should not be used if following the PCR-free protocol.

  • What does Revvity do to assure quality?

    Please refer to our dedicated page describing our quality assurance philosophy.

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