NEXTFLEX Cell Free DNA-Seq Library Prep Kit 2.0_48 RXN

RevvitySKU: REV-NOVA-5150-02



  • Designed for low sample input- 10 ng of input cfDNA required
  • Accelerated workflow with minimal hands-on time
  • Flexible adapter barcode options- up to 384 single index or unique dual index barcodes are available
  • Compatible with Illumina® sequencing platforms
  • Protocols available for automation on the Revvity® Sciclone® and Zephyr® G3 NGS workstations

 Prepare Libraries in Only 2 Hours

The NEXTFLEX® Cell Free DNA-Seq Kit 2.0 can produce libraries from 10 ng of cell free DNA for circulating tumor DNA or cell free fetal DNA (cffDNA) analysis, in two hours. This low-input library preparation kit delivers high coverage quality and reduced bias for Illumina® sequencing applications. The NEXTFLEX® Cell Free DNA-Seq Kit 2.0 can be used to prepare single, paired-end and multiplexed DNA libraries for sequencing using Illumina®platforms. The availability of up to 384 single-index single index and unique dual index barcodes facilitates high-throughput applications without compromising data integrity.

We recommend using the NextPrep-Mag cfDNA Automated Isolation Kit for isolation of circulating cfDNA from blood plasma prior to library prep with the NEXTFLEX® Cell Free DNA-Seq Kit 2.0.

 Reduced Bias and Coverage Gaps

NEXTFLEX® Cell Free DNA-Seq Kit 2.0 exhibits minimal GC bias and produces uniform coverage of difficult to sequence genome regions. This kit also offers robust ligation efficiency, resulting in greater library diversity and a larger number of unique sequencing reads. These optimizations enable investigators to obtain more information from each sequencing run.

Flexible Multiplexing Options

The NEXTFLEX® Cell Free DNA-Seq Library Prep Kit 2.0 for Illumina® sequencing is designed to be used with the NEXTFLEX® Unique Dual Index barcodes, NEXTFLEX® DNA barcodes, NEXTFLEX-96 DNA barcodes and NEXTFLEX-HT barcodes. These barcodes are available in sets of 6, 12, 24, 48, 96 and 384 unique indexed adapters. All of these NEXTFLEX® Adapters are long, annealed adapters that offer an improved multiplexing workflow and flexible setup.


  • NEXTFLEX® cfDNA Enrichment Beads
  • 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 (50 μM)
  • Nuclease-free Water
  • NEXTFLEX® Resuspension Buffer
  • NEXTFLEX® Cleanup Beads 2.0 1.5 mL


  •  ~1 ng of Cell-Free DNA in up to 32µL nuclease-free water.
  • NEXTFLEX® DNA Barcodes- 6 / 12 / 24 / 48 (Cat # 514101, 514102, 514103, 514104) or NEXTFLEX-96 DNA Barcodes (Cat # 514106)
  • Ethanol 100% (room temperature)
  • Ethanol 80% (room temperature)
  • 96 well PCR Plate Non-skirted (Phenix Research, Cat # MPS-499) or similar
  • 96 well Library Storage and Pooling Plate (Fisher Scientific, Cat # AB-0765) or similar
  • Adhesive PCR Plate Seal (BioRad, Cat # MSB1001)
  • Magnetic Stand -96 (Ambion, Cat # AM10027) / or / similar
  • Thermocycler
  • 2, 10, 20, 200 and 1000µL pipettes / multichannel pipettes
  • Nuclease-free barrier pipette tips
  • Vortex

Kit Specifications:

The NEXTFLEX® Cell Free DNA-Seq Kit 2.0 contains enough material to prepare 8 or 48 DNA samples for Illumina® compatible sequencing when bundled with the NEXTFLEX® NGS barcodes. The shelf life of all reagents is 6 months when stored properly. The Resuspension Buffer and Nuclease-free Water can be stored at room temperature. The NEXTFLEX® cfDNA Enrichment Beads and Cleanup Beads 2.0 should be stored at 4°C, and all other components should be stored at -20°C.


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