新一代高通量基因测序NGS技术和临床应用-AuroraBiomed.PDFVIP

Clinical Application of Ampliseq-Targeted Sequencing Ardent BioMed Guangdong Inc. （广东国盛医学科技股份有限公司） David Deng （鄧杏飛） Topics  Brief NGS Introduction  Methods of Targeted NGS Library Preparation  Clinical Application: Examples of Ampliseq Panel Design 2 Precision Medicine - Requires Precision Detection First! 3 Basic NGS Procedures DNA Extraction/ Library Sequencing Data Analysis Fragmentation Preparation Adapters Ligation • Random(Physical ） Seq. by Synthesis • Endonuclease Amplification Seq. by Semiconductor Chip DNA Template Preparation 4 （Emulsion PCR ） Single Molecular Seq. Types of NGS By Sequencing Contents  Whole Genome Sequencing （WGS ）  Whole Exome Sequencing （WES ）  Targeted Sequencing By Sample Types and Purposes  RNA Seq.  DNA Seq.  Single Cell Seq.  DNA Methylation Seq. 5 Factors Affecting NGS  Quality of Nucleic Acid Extraction  DNA Integrate Quality ：Sample preservation and Extraction  False positive or false negative results – Oxidation-damaged DNA （published results ）  DNA Fragmentation  Appropriate size and even DNA fragments: Key factors  Physical ；Enzymatic or Chemical  Library Preparation – Targeted Amplification  Routine clinical application of NGS: targeted sequencing of known mutations  Selective amplification/Capturing DNA fragme