遗传学：SOLiD_产品介绍.pptVIP

SOLiD? System OverviewABI 新一代高通量基因分析系统 Applied Biosystems SOLiD? System – 概况 SOLiD? System 为高通量并行测序技术平台,采用结合在磁珠上的克隆扩增的单分子DNA 片段为测序模板 测序通量 : Up to 3 GB of data/run (1.5GB per slide)? 2 slide system 测序读长 : 35bp fragment, 2x25bp in mate pair SOLiD – 工作流程 1. Prepare fragment or mate-paired library from starting material (DNA or RNA)? 2. Amplify library onto beads using emulsion PCR 3. Randomly deposit bead clones onto slide surface Roche –Pico Titre Plate – Limited number Illumina – Flow Cell with fixed # of sites 4. Sequence clones by ligation-based sequencing Roche and Illumina 均采用polymerase Workflow 油包水PCR: Emulsion PCR (ii)? emulsion PCR Individual Bead emulsion PCR Individual Bead Beads collected following emulsion PCR: 纯化 Enrichment P2-hybridization, Pre- and post-enrichment 测序芯片 Sequencing Array  Properties of the Probes Spatial separation among dye, ligation cleavage sites SOLiD Chemistry System 4-color ligationLigation reaction SOLiD Chemistry System 4-color ligation Ligation reaction SOLiD Chemistry System 4-color ligation De-Phosphorylation SOLiD Chemistry System 4-color ligation Visualization SOLiD Chemistry System 4-color ligation Cleavage SOLiD Chemistry System 4-color ligation Ligation (2nd cycle)? SOLiD Chemistry System 4-color ligation Visualization (2nd cycle)? SOLiD Chemistry System 4-color ligation Cleavage (2nd cycle)? SOLiD Chemistry System 4-color ligation interrogates every 5th base SOLiD Chemistry System 4-color ligation Reset SOLiD Chemistry System 4-color ligation  (1st cycle after reset)? SOLiD Chemistry System 4-color ligation  (1st cycle after reset)? SOLiD Chemistry System 4-color ligation  (2nd Round)? Sequential rounds of sequencingMultiple cycles per round 2 base encoding 5 Major Breakthroughs of SOLiD? System Applied Biosystems SOLiD? System - Overview Key differentiators – SOLiD Solexa Roche Bases/Run 3GB 1GB 0.1GB Cost/Run ~$3000 ~$3000 ~$10000+ Lg. Mat