Hieff NGS™ OnePot Pro DNA Library Prep Kit V4 is is an advanced enzymatic DNA library preparation kit designed for compatibility with both Illumina™ and MGI™ next-generation sequencing(NGS) platforms. By utilizing high-efficiency enzymatic fragmentation, this kit eliminates the need for traditional mechanical shearing methods such as sonication, streamlining the workflow and minimizing hands-on time. The kit integrates DNA fragmentation and end-repair into a single-step reaction, significantly reducing overall library preparation time and cost. It is suitable for a broad range of sample types, including plant genomes, animal genomes, microbial genomes, and more, with a flexible DNA input range from 1 ng to 1 μg. The optimized enzymatic fragmentation ensures consistent and uniform library fragment sizes across different species, minimizing variation and enhancing sequencing performance. The prepared libraries are fully compatible with Illumina™ or MGI™ adapters and primers, enabling seamless downstream sequencing on respective platforms. Features Broad Sample Compatibility: Supports a wide range of input DNA from 100 pg to 1 µg, covering all common sample types, such as human, animal and plant DNA. High Fragment Recovery: Post-fragmentation recovery rate exceeds 60%. Excellent Library Uniformity: Consistent library distribution across different species under the same preparation conditions. Strong Inhibitor Tolerance: High success rate of direct library preparation from complex samples containing inhibitors. Automation-Friendly: Single-use reagents remain stable for 4 weeks at 4°C or 25°C, facilitating automation. Components Components No. Name 12972ES08 12972ES24 12972ES96 12972-A Smearase™ Buffer 4.0 80 μL 240 μL 960 μL 12972-B Smearase™ Enzyme 4.0 80 μL 240 μL 960 μL 12972-C Ligation Enhancer 4.0 240 μL 720 μL 3×960 μL 12972-D Rapid DNA Ligase 4.0 80 μL 240 μL 2×480 μL 12972-E 2×Ultima HF Amplification Mix 200 μL 600 μL 3×800 μL Storage This product should be stored at -25~-15℃ for one year. Application DNA Library Preparation; Whole Genome Sequencing(WGS); Whole Exome or other Targeted Capture Sequencing; Amplicon Sequencing; Immunoprecipitation Sequencing; Metagenomic Sequencing; Figures 1.Flexible and High-Efficiency Fragmentation Figure 1. Fragment size distribution of 250 ng human gDNA using the 12972 fragmentation module. Human gDNA (250 ng) was fragmented at 30°C or 35°C for 10, 15, 20, 25, 30, or 40 minutes. The distribution of recovered insert sizes is shown, demonstrating stable and uniform fragmentation across the 10–40 minute range. 2.Excellent Buffer Tolerance Figure 2. Fragmentation performance of 12972ES in different reaction buffers(H2O (pH 7.0), 1×TE (pH 7.5–8.0), 1×AE (pH 8.3), and 1×EB (pH 8.5)). Fragmentation reactions were performed using 12972ES with 500 ng of 293 gDNA at 30°C for 20 minutes in four different buffers. When 10 μL of each buffer was used with DEPC-treated water, the fragment sizes were similar to those in pure DEPC water. Using AE or EB as the full reaction buffer resulted in slightly larger fragments. 3.Broad Species Compatibility Figure 3. Library uniformity testing across different species using 12972 under identical fragmentation conditions. Different DNA samples were fragmented with 12972 under identical conditions. The libraries showed consistent sizes (~450 bp / ~320 bp) with good uniformity across species. 4.Excellent Sequencing Data Performance Figure 4. Performance of 10X WGS Data on Human and Bovine Samples 12972 demonstrates higher gene coverage compared to Supplier W*. Both false chimeric rate and hairpin artifacts are lower or comparable to those of Supplier W*. 5.Strong Stability Figure 5. Library yield stability after storage at 4°C and 25°C for 4 weeks. No decrease in library yield was observed after 4 weeks of storage at 4°C and 25°C, highlighting the kit’s suitability for automation. Documents: Safety Data Sheet 12972_MSDS_HB250620_EN.PDF Manuals 12972_Manual_HB20260401.pdf Protocol 12972_PCR-free Library Prep Protocol _Ver.EN20260623 Related Blog: Enzymatic Fragmentation DNA Library Prep Kits: Empowering Multi-Disciplinary Research Applications
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