NEXTFLEX Rapid DNA-Seq Kit 2.0

Higher WGS library yield & conversion across wide input range

The NEXTFLEX Rapid DNA-Seq 2.0 consistently converts a high percentage of mechanically-sheared fragments into adapter-ligated molecules, delivering up to 2 × more final library (nM) than a leading mechanical-ligation kit, even at 1 ng DNA input. The higher conversion translates to a larger pool of unique reads and more uniform coverage in GC-rich regions (≤ 3 % GC-slope).

Figure 1. Superior yield and conversion with NEXTFLEX Rapid DNA-Seq 2.0.
(A) Library yield (nM) from 250 ng and 1 ng inputs of human genomic DNA and E. coli DNA; libraries were amplified with identical PCR-cycle counts. NEXTFLEX Rapid DNA-Seq 2.0 (blue) produces substantially higher yields than Competitor K (purple), especially at 1 ng.
(B) Normalised library-conversion rate (ratio of qPCR-quantified library molecules to input fragment count) for 250 ng human DNA shows.

Reduced GC bias delivers more even genome coverage

GC-coverage analysis shows that NEXTFLEX Rapid DNA-Seq 2.0 preserves uniform read depth from 20% to 80% GC, whereas other mechanical-ligation kits lose reads in GC-rich windows. This uniformity translates to near-complete genome coverage at standard sequencing depths, allowing researchers to call SNPs and assemble contigs with confidence, even in regions that are typically under-represented.

Figure 2: GC-coverage plots show uniform read depth with NEXTFLEX Rapid DNA-Seq 2.0 versus coverage loss in competitor libraries

Greater usable genome coverage at standard sequencing depths

When libraries are sequenced to typical whole-genome depths, NEXTFLEX Rapid DNA-Seq 2.0 delivers a larger proportion of the genome above key coverage thresholds (1X, 5X, 10X) than a leading mechanical-ligation kit.  This higher usable coverage, observed in both yeast and E. coli datasets, reduces the need for additional sequencing reads and improves confidence in downstream SNP discovery, structural-variant calling, and de-novo assembly projects.

Figure 3: At equivalent read depths, libraries prepared with NEXTFLEX Rapid DNA-Seq 2.0 (blue) cover a larger fraction of the yeast and E. coli genomes at ≥ 1X, ≥ 5X and ≥ 10X thresholds than those prepared with Competitor K (purple).  The improved coverage reduces re-sequencing costs and enhances variant-calling accuracy in whole-genome studies.

Flexible Multiplexing Options

The NEXTFLEX Rapid DNA-Seq Kit 2.0 supports high-throughput multiplexing with the full line of NEXTFLEX Adapters  offering:

• Up to 1,536 unique dual-index pairs (8- or 10-bp) to eliminate index hopping and enable large WGS or capture projects
• Single-index and paired-end compatibility for legacy workflows
• UDI-UMI plates (96 pairs) that add unique molecular identifiers for accurate low-frequency variant detection and duplex sequencing
• Color-balanced index sets optimized for Illumina® patterned flow-cells and Element AVITI™ chemistry

Automation Compatibility

The NEXTFLEX Rapid DNA-seq Kit 2.0 is compatible with multiple liquid handlers, including the Revvity Sciclone G3 NGSx and Zephyr G3 NGS workstations.

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