DESIGNED TO PREPARE MULTIPLEXED LIBRARIES FOR SINGLE OR PAIRED-END SEQUENCING USING ILLUMINA® PLATFORMS

NEXTFLEX® Rapid DNA-Seq Kit 2.0 for Illumina® Platforms

  • Complete library prep solution, including size selection beads
  • Bundle kits available with library prep reagents and barcodes
  • Up to 384 barcodes available upon request
  • Convenient and cost-effective package in a 8, 48, or 96 reaction format
  • Simplified workflow
  • Automation compatible with liquid handlers, including the PerkinElmer Sciclone® G3 NGS/NGSx workstations and Zephyr® G3 NGS workstation
  • Robust genome coverage
  • Reliable performance with GC-bias and sequencing bias mitigation
  • Low input requirement for genomic DNA down to 1 ng
  • For FFPE samples, use the NEXTFLEX® Rapid XP DNA-Seq Kit
  • Functionally validated with Illumina® sequencing platforms
Download Manual
For research use only. Not for use in diagnostic procedures.

  • NOVA-5188-01

       $198

    (NO BARCODES)
    8 RXNS
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       $1,160

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    48 RXNS
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  • NOVA-5188-03

       $2,252

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    96 RXNS
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  • NOVA-5188-11

       $237

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  • NOVA-5188-12

       $1,380

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BUNDLE

  • NOVA-5188-13

       $2,719

    BARCODES 1-96
    96 RXNS
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Ligation of Adapters to SARS-CoV-2 Amplicon Pools

After generating amplicon pools with SARS-CoV-2 specific primers, you need to add adapters to them to create amplicon libraries for sequencing. While there are many options available, the ability to multiplex up to 1,536 samples simultaneously greatly reduces the both the cost and sequencing time when compared to options that can only multiplex 384 samples at a time.

If your SARS-CoV-2 primer sets for amplicon sequencing generate short tiled amplicons which don’t require fragmentation, we recommend using the NEXTFLEX® Rapid DNA-seq kit 2.0 with the NEXTFLEX® UDI barcodes. This solution allows researchers to multiplex up to 1,536 samples per run, increasing throughput and reducing the cost and time required for sequencing.

If you are using long cDNA which requires fragmentation prior to sequencing, use the NEXTFLEX® Rapid XP DNA-seq kit.

Rapid DNA Library Prep for Sequencing on Illumina® Platforms

The NEXTFLEX® rapid DNA-seq 2.0 kit is designed to prepare multiplexed libraries for single or paired-end sequencing using Illumina® platforms. The enhanced NEXTFLEX® kit simplifies workflow by using master mixed reagents and magnetic bead based cleanup, reducing pipetting and eliminating time-consuming steps in library preparation. The NEXTFLEX® rapid DNA-seq kit 2.0 produces library preps with a larger number of unique sequencing reads, allowing for robust genome coverage in challenging GC-rich sequences that can introduce bias.

The NEXTFLEX® rapid DNA-seq 2.0 kit bundle offers all the advantages and reagents of the NEXTFLEX Rapid DNA-seq kit 2.0 and includes  NEXTFLEX-HT™ barcodes (8, 48, or 96 reaction size kits) supplied in a single-use format. This allows the lab to tie all the necessary components of the library prep workflow to a single lot for ease of purchasing and tracking of consumables.

Figure 1: Libraries prepared with NEXTflex® rapid DNA-Seq kit 2.0 show higher yield than libraries prepared with Competitor K's kit using the same number of PCR cycles from a 250 ng and 1 ng input (A) and robust conversion rate (B).

Figure 1: Libraries prepared with NEXTFLEX® rapid DNA-Seq kit 2.0 show higher yield than libraries prepared with Competitor K’s kit using the same number of PCR cycles from a 250 ng and 1 ng input (A) and robust conversion rate (B).

Flexible Multiplexing Options

The NEXTFLEX® adapters are long, annealed adapters containing indexed sequences, which offer an improved multiplexing workflow and flexible experimental setups catered to individual needs. Our barcode portfolio encompass options for single or paired-end multiplex sequencing, and the NEXTFLEX® rapid DNA-seq kit 2.0 is compatible with the NEXTFLEX® DNA Barcodes and NEXTFLEX-96™ DNA Barcodes, NEXTFLEX® ChIP-Seq Barcodes, and NEXTFLEX-96™ ChIP-Seq Barcodes. The NEXTFLEX® rapid DNA-seq kit 2.0 reagents are also compatible with the NEXTFLEX® Unique Dual Index Barcodes  for up to 192 sample multiplexing.

The NEXTFLEX® rapid DNA-seq kit 2.0 bundles come provided with the NEXTFLEX-HT™ Barcodes.

For more information about barcode options compatible with the NEXTFLEX® rapid DNA-seq kit 2.0, please contact [email protected].

Automation Compatibility

The NEXTflex® rapid DNA-seq kit 2.0 is compatible with multiple liquid handlers, including the PerkinElmer Sciclone® G3 NGS and NGSx workstations. For more information, or to obtain a program, contact [email protected].

For larger volume requirements, customization and bulk packaging is available. For increased flexibility, individual reagents (non-master mixed) are also available. Please contact [email protected] for further information.

Tech Tips – When to Size Select

Size selection is a critical issue in NGS library preparation. Our blog post, Tech Tips – When to Size Select, addresses a number of factors to consider before determining a size selection strategy.

KIT CONTENTS

  • NEXTFLEX® End Repair & Adenylation Buffer Mix 2.0
  • NEXTFLEX® End Repair & Adenylation Enzyme Mix 2.0
  • NEXTFLEX® Ligase Buffer Mix 2.0
  • NEXTFLEX® Ligase Enzyme 2.0
  • NEXTFLEX® PCR Master Mix 2.0
  • NEXTFLEX® Primer Mix 2.0
  • NEXTFLEX® Cleanup Beads 2.0
  • Nuclease-free Water
  • Resuspension Buffer

REQUIRED MATERIALS NOT PROVIDED

  • 1 ng – 1 µg of fragmented genomic DNA in up to 32 µL nuclease-free water.
  • NEXTflex® barcodes if purchasing a non-bundled kit (includes NEXTflex-HT™ barcodes)
  • Ethanol 100% (room temperature)
  • Ethanol 80% (room temperature)
  • Covaris System (S2, E210) or other method for DNA fragmentation
  • 96 well PCR Plate Non-skirted (Phenix Research, Cat # MPS-499) or similar
  • 96 well Library Storage and Pooling Plate (Thermo Fisher Scientific, Cat # AB-0765) or similar
  • Adhesive PCR Plate Seal (Bio-Rad, Cat # MSB1001)
  • Magnetic Stand -96 (Thermo Fisher Scientific, Cat # AM10027) or similar
  • Heat block
  • Thermocycler
  • 2, 10, 20, 200 and 1000 µL pipettes / multichannel pipettes
  • Nuclease-free barrier pipette tips
  • Microcentrifuge
  • 1.5 mL nuclease-free microcentrifuge tubes
  • Vortex

NEXTFLEX Rapid DNA-Seq Kit 2.0 Protocol

Additional Data between NEXTFLEX® Rapid DNA-Seq 2.0 and Competitor K’s Kit Libraries

Rapid DNA-Seq Kit 2.0 data
Figure 2: Libraries prepared with NEXTFLEX® Rapid DNA-seq 2.0 shows less GC bias than libraries prepared with Competitor K’s kit.

Rapid DNA-Seq Kit 2.0 data
Figure 3: Libraries prepared with NEXTFLEX® Rapid DNA-seq 2.0 shows higher genome coverage than libraries prepared with Competitor K’s kit.

The NEXTFLEX®  rapid DNA-seq 2.0 kits contain library prep reagents, including cleanup beads, to prepare 8, 48, or 96 genomic DNA samples for Illumina® sequencing. The bundled solution also includes the corresponding number of unique NEXTFLEX-HT™ barcodes per reaction. The shelf life of all reagents is 6 months when stored properly at -20°C. This kit is shipped on dry ice.

Adapters for use with the NEXTFLEX® Rapid DNA-seq Kit 2.0 for Illumina® sequencing (Note: the bundles come with the NEXTFLEX-HT™ Barcodes)

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