Hieff NGS™ OnePot Evo DNA Library Prep Kit is a next-generation enzymatic library preparation kit compatible with both Illumina and MGI high-throughput sequencing platforms. Compared to traditional library construction methods, this kit replaces time-consuming and instrument-dependent sonication with a high-quality, low-bias fragmentation enzyme. It uniquely integrates the fragmentation, end repair, and dA-tailing steps into a single reaction, significantly reducing hands-on time, cost, and workflow complexity. The ligation module features pre-mixed enzyme and buffer for streamlined operation, making the kit highly suitable for automation. Building upon the previous version, this updated formulation further reduces GC bias, enhances end-repair and dA-tailing efficiency, and improves reagent stability. It also incorporates a newly optimized ligase for superior adapter ligation performance. The kit is fully compatible with standard Illumina or MGI adapters and primers. Ideal for whole-genome sequencing of animals, plants, microbes, and challenging clinical samples. Features Wide input range: Compatible with 5 ng–1 µg of genomic DNA, full-length cDNA, or other double-stranded DNA High-quality fragmentation enzyme: Enables random dsDNA cleavage with minimal sequence bias, integrating fragmentation, end repair, and dA-tailing in a single step High-fidelity, high-efficiency PCR enzyme: Maximizes library yield and complexity while preserving sequence accuracy FFPE-compatible: Delivers robust performance with degraded or formalin-fixed DNA samples Rigorous quality control: Each production batch undergoes stringent performance validation and stability testing Specifications Cat.No. 12193ES08 / 12193ES24 / 12193ES96 Size 8 T / 24 T / 96 T Components Components No. Name 12193ES08 12193ES24 12193ES96 12193-A Smearase™ Buffer 40 μL 120 μL 480 μL 12193-B Smearase™ Enzyme 80 μL 240 μL 960 μL 12193-C Rapid Ligase Master Mix 280 μL 840 μL 4×840 μL 12193-D 2× Ultima HF Amplification Mix 200 μL 600 μL 3×800 μL [Note]: The kit components are compatible with both Illumina &MGI sequencing platform, if the complete adapter was used, Hieff NGS Primer Mix (Yeasen Cat#12190 or Cat#12191) in needed. Application DNA library Preparation; Whole genome sequencing(WGS); whole exome or other targeted capture sequencing; amplicon sequencing; immunoprecipitation sequencing; metagenomic sequencing; Figures 1. Flexible & High-Efficiency Fragmentation Figure 1. Fragmentation time gradient using 200 ng human gDNA. Digestion was performed at 3, 5, 7, 10, 12, 15, 17, and 20 min, yielding target fragments of 200–1,000 bp. 2. Broad Input Compatibility and High Yield. Figure 2. Library construction performance for gDNA and FFPE samples over varied input quantities. Following enzymatic fragmentation at 37 °C for 10 min, high-yield libraries are consistently generated. The recommended PCR cycles vary by sample type and input amount: for gDNA, 10 cycles at 5 ng, 6 cycles at 50 ng, and 3 cycles at 500 ng–1 μg; for FFPE samples, 7 cycles at 50–100 ng and 6 cycles at 200–300 ng. 3. Exceptional Capture Performance (WES). (A). WES Performance (NA12878 Standard). Figure 3. Superior Capture Data Quality.(A). WES Performance (NA12878 Standard). Compared to Supplier A, 12193 exhibits significantly higher properly paired read rates and target coverage, alongside a markedly lower proportion of pairs mapped to different chromosomes, indicating superior library integrity. (B). Robust Targeted Capture (FFPE Samples). Figure 4. Superior Capture Data Quality.(B). Targeted Capture Performance (FFPE Samples). Robust Targeted Capture (FFPE Samples). Using the same 0.2M Panel, 200 ng input, and 5 PCR cycles, 12193 delivers the highest library yield and best-in-class Map Ratio, Dup, Captured Reads, and Mean Depth. Although ranking second for Chimeric and Hairpin Artifacts, 12193 provides a superior overall data quality and yield profile compared to Supplier V*. 4. Customer Case Figure 5. Performance Comparison of FFPE Libraries. The 12193 workflow delivers robust and reproducible performance. Both samples exhibit low duplication rates (~33%, well below the 40% threshold) and exceptional mapping rates (>99.9%). The libraries achieve high uniformity (~99.7%), deep mean coverage (>8,700×), and a high proportion of usable on-target bases (24.3%). Furthermore, 100% SNV concordance confirms the high accuracy of the assay.
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