The Hieff DNA Library Prep Kit for ONT is a DNA library preparation kit developed for the third-generation Nanopore sequencing platform. This kit has undergone targeted modifications and optimizations to its enzymes and buffer system, significantly improving the ligation efficiency of long fragments. Additionally, after end repair and A-tailing, no purification is required, allowing direct Barcode ligation, making the experimental workflow more convenient. Applications DNA Library Construction Third-Generation Library Construction TGS Library Construction Nanopore Sequencing ONT Sequencing Product Advantages Ease of Operation: End repair and A-tailing completed in one step, no purification required, allowing direct ligation with Barcode adapters Wide Compatibility: Compatible with various sample types including animals, plants, and microorganisms; supports both gDNA and amplicon samples Excellent Data Performance: High library yield and superior sequencing data quality Components No. Name 13301ES08 13301ES24 13301ES96 8 T 24 T 96 T 13301-A Endprep Buffer 56 μL 168 μL 672 μL 13301-B Endprep Enzyme 24 μL 72 μL 288 μL 13301-C Rapid Ligase Master Mix 280 μL 840 μL 4×840 μL 13301-D Rapid Ligation Reaction Buffer（5×） 160 μL 480 μL 2×960 μL 13301-E Rapid T4 DNA Ligase 40 μL 120 μL 480 μL 13301-F Elution Buffer 160 μL 480 μL 2×960 μL Precautions 1. The washing reagents used for purification after Native Barcode ligation and Native Adapter purification are different. Please ensure the correct reagents are used for each purification step. 2. To ensure successful library construction, it is recommended to verify the fragment length and integrity of the input DNA (validation methods include capillary electrophoresis or other equivalent methods). DNA quality (integrity, fragmentation), purity (e.g., chemical contamination, RNA residuals), and excessive or insufficient usage may affect library yield. 3. Please prepare all necessary materials in advance according to the provided guidelines to avoid issues that could prevent proper execution of the experiment, leading to sample or reagent waste. 4. When preparing libraries for multiple samples, please record the Native Barcode number corresponding to each sample. Workflow Case Study Template Input DNA Recovery Data Volume Reads mean length Reads max length Reads N50 length Human gDNA (4-20 K) 1 μg 96% 15 G 9136.1715 113518 12633 1.7 K 500 ng 65% 0.33G 1791 23493 1771 3 K 500 ng 75% 0.31G 2557 26165 3005 1-16 K 1 μg 34.5% 2.89G 4189 41457 5893 Plasmid 1 300 ng ＞60% 0.96 G 2413 23637 2658 Plasmid 2 300 ng ＞60% 0.83 G 2129 35855 2808
