单细胞组学及其应用.pptxVIP

Single Cell Omics and Its Application Content OverviewNext Generation Single Cell AnalysisSingle cell DNA sequencing and its applicationSingle cell RNA sequencing and its applicationApplications on CTC 3The front-end and back-end of single cell analysisCell isolationOmics techMicrofuidics Microscopic capture/micromanipulation4What is Single Cell Omics?Front-EndDisadvantage:Throughput;Resolution;Advantage:Easy to access;Easy to use; Fluorescence Activated Cell Sorting (FACS)5What is Single Cell Omics?Front-EndDisadvantage: Cell damage;Fluoresce affect amplificationAdvantage: High throughput;Specific markers; Laser capture microdissection/laser microdissection6What is Single Cell Omics?Front-EndDisadvantage:Cell damage and information loss;Throughput;Advantage:Micro-environment;Specific markers and specific region Microfluidics:7What is Single Cell Omics?Front-EndDisadvantage:User friendly? Modulated?Advantage:High throughput; Automatic 8What is Single Cell Omics? What is Single Cell Omics?BridgingFluidigm C1 SystemMIRALCS System Single cell DNA sequencing and its application Single Cell Genomics SamplesSingle CellNGSCell isolation WGASequencing the Single Cell Genome by Next Generation Sequencing (NGS)Library Preparation Enough Amount of DNABioinformatics Analysis12Single Cell Genomics Research 13Whole Genome Amplification (WGA)(Zong,2012)Single Cell GenomicsMultiple Displacement Amplification (MDA)Multiple Annealing and Looping Based Amplification Cycles (MALBAC) No obvious genome wide coverage limitation by single cell sequencing14Methodology of Single Cell SequencingGC content does impact the even distribution of WGA data. Acceptable sequencing error rateFN FDR sites do not show specific base type bias beyond mutationsAmplification failure genes do not show preference on different biological processes Sample DescriptionTumor type Single cell number tumor cell #/control cell #(sequencing available number)GenderDescriptionEssential thrombocytosis (ET)81/9(58/8)