新一代测序 (NGS) 通过实现高通量、经济高效的基因组和转录组测序，加速了包括癌症在内的复杂疾病的个性化医疗，彻底改变了基因组研究。全基因组/转录组测序 (WGS/WTS) 提供全面的见解，而靶向测序更具成本效益和敏感性。与由于高速和成本效益而仍然占据主导地位的短读长测序相比，长读长测序可以克服比对限制并更好地区分来自替代转录本或重复区域的相似序列。混合测序结合了不同技术的最佳优势，可以更全面地了解基因组/转录组变异。了解每种技术的优点和局限性对于将尖端技术转化为临床应用至关重要。在这项研究中，我们使用各种靶向 DNA 和 RNA 组合以及短读长和长读长平台上的整个转录组对参考样本的 DNA 和 RNA 文库进行了测序。该研究设计能够对测序技术、靶向方案和文库制备方法进行全面分析。我们扩展的分析图谱为评估当前测序技术建立了参考点，促进基因组研究和精准医学的明智决策。© 2024。这是美国政府的作品，在美国不受版权保护；外国版权保护可能适用。

Next-generation sequencing (NGS) has revolutionized genomic research by enabling high-throughput, cost-effective genome and transcriptome sequencing accelerating personalized medicine for complex diseases, including cancer. Whole genome/transcriptome sequencing (WGS/WTS) provides comprehensive insights, while targeted sequencing is more cost-effective and sensitive. In comparison to short-read sequencing, which still dominates the field due to high speed and cost-effectiveness, long-read sequencing can overcome alignment limitations and better discriminate similar sequences from alternative transcripts or repetitive regions. Hybrid sequencing combines the best strengths of different technologies for a more comprehensive view of genomic/transcriptomic variations. Understanding each technology's strengths and limitations is critical for translating cutting-edge technologies into clinical applications. In this study, we sequenced DNA and RNA libraries of reference samples using various targeted DNA and RNA panels and the whole transcriptome on both short-read and long-read platforms. This study design enables a comprehensive analysis of sequencing technologies, targeting protocols, and library preparation methods. Our expanded profiling landscape establishes a reference point for assessing current sequencing technologies, facilitating informed decision-making in genomic research and precision medicine.© 2024. This is a U.S. Government work and not under copyright protection in the US; foreign copyright protection may apply.