GeneChip概览.pptVIP

Genomewide Associations Reported through March 2010. Examples of Previously Unsuspected Associations between Certain Conditions and Genes and the Related Metabolic Function or Pathway, According to Genomewide Association Studies. Examples of Loci Shared by Conditions or Traits Previously Thought to Be Unrelated, According to Genomewide Association Studies. 基因芯片的将来 未来会被高通量测序技术取代 ChIP-chip 不如ChIP-Seq 但目前几年内芯片不会退出舞台 NGS成本太高 NGS技术不会像芯片技术那样成熟 芯片技术有很多已有数据可以比较 芯片技术已广泛用于NGS样品制备 Exon capture arrays Thank you! zhizheng10@ * * 该文章可以网上下载，也有印刷的资料 NATURE BIOTECHNOLOGY是级别相当高的杂志 目前为止，所有ILLUMINA平台上发表的文章都在高级别的杂志上.最为最新的芯片技术，我们的发展令业内震惊，也在越来越多实验室得到应用和肯定 * Actually a number of clustering methods have been proposed. I’ll go through some representative types in the following slides. * * Here is a cartoon to show how the bead assay is run * Need not only detect assay signal, also decode the beads * Because beads mixed as suspension,… Beads overcome many limitations of planar array---platform used in clinical diagnostic Still the bead array described so far , the hyb protocol is similar to mA. So sensitivity is still low 芯片结果难以重复的原因 使用的芯片平台不同，探针不同 样品制备步骤复杂，结果难以重复 数据分析方法不尽相同 样品数目不够造成关联为假阳性 怎样验证芯片结果的确代表Biomarkers？ 在大量独立样本中进行验证（validation） 使用高样本通量技术，检测几百个样品，每样品检测几十个基因表达 开发几乎无需样品制备步骤的基因检测技术 目前广泛使用的验证技术：qRT-PCR ？？？ 对少量candidate genes 小规模验证， qRT-PCR 可以 Bead Technology 1-100 plex assays Small sample volume 1,000 samples /day Reproducible results 悬浮芯片（Liquid Arrays :一种适用于验证芯片结果的平台技术 Microparticles For Multiplexed Assays One run, up to 96 samples, Each sample, up to 100 genes tested. Multiplexed, High Throughput Assay Detection in Bead Assays Multiplex Analysis on Luminex Beads “Liquid Array” vs. Planar Array Hyb kinetics: near solution phase solid phase Speed fast slow Statistical sampling N 100 N 1 Flexibility Easy to add genes Not flexible Sample throughput 96 or 384 1-10 Gene number 1-100 1000-20,000 一种适用临床的基因检测新技术 在细胞样品甚至血液中直接检测RNA含量 不需要样品提纯,RNA抽取,标记或扩增,不用任何酶反应 在96