VAHTS AmpSeq Library Prep Kit V3 is based on ultra-multiplex PCR, introducing several core technologies such as end primer digestion, to construct amplicon libraries. The entire library preparation workflow of this kit takes place in a single tube, the sequencing library can be obtained after purification. It takes as little as 5 h from amplification to the generated library, including less than 1.5 h of manual operation. The initial template for this kit is 1 - 100 ng. It is compatible with genomic DNA, FFPE samples, cfDNA, etc. This kit is intended for both the Illumina and Ion Torrent high-throughput sequencing platforms. And corresponding adapters (Vazyme #NA111/ NA121) are also provided. It is compatible with panels including Ion AmpliSeq Series Panel and AmpliSeq for Illumina Series Panel, as well as the corresponding individually customized panels. Compared to the previous generation (Vazyme #NA201), the library preparation and sequencing quality have been greatly improved. The optimized multiplex amplification module has good amplification performance for different input amounts and customized panels. The upgraded digestion module can effectively solve the problem of PCR products contamination and improve data utilization. The newest ligation module increases the effective library ratio. Comprehensive product upgrade ensures the high coverage and high uniformity of the amplicon library. With this kit, the sequencing results are more stable and reliable, helping researchers and testing personnel to complete high quality library preparation quickly and easily. All the reagents provided in the kit have undergone rigorous quality control and functional testing, to ensure the optimal stability and repeatability of library preparation.