临床基因组学：第五章 染色体微阵列分析.ppt

GeneChip? Fluidic Station 450Dx v.2 GeneChip? 3000Dx v.2 with AutoLoader 微阵列技术相关网络资源 广州国际生物岛螺旋三路10号 做 最 优 秀 的 第 三 方 检 测 机 构 TO BE THE BEST INDEPENDENT LABORATORIES 模板来自于 / * 三倍体 三倍体发生机制是由于受精过程中生殖细胞分裂异常导致，主要有双雄受精和双雌受精。表型与基因组印迹相关。 母源性三倍体多为早期自然流产，胎儿严重发育障碍，胎盘细小不发育但无葡萄样变。 父源性三倍体常表现为局部性葡萄胎样大胎盘，胎儿小，宫内发育迟缓，先天性心脏病，并指等。三倍体胎儿都不能成活出生，大部分在10~20孕周流产。 嵌合体 mosaicism CMA 技术的实验流程 GeneChip? Hybridization Oven 645 GeneChip? Fluidic Station 450Dx v.2 GeneChip? 3000Dx v.2 with AutoLoader Agilent SureScan Dx Microarray Scanner 实验流程 实验流程 酶切消化 Digestion 酶切消化 Digestion 酶切消化 Digestion 连接 Ligation 连接产物稀释 25ul产物+75ul水 PCR扩增 PCR扩增 2% TBE agarose gel at 5 v/cm for 45 minutes. Average product distribution is between ~150 to 2000 bp 磁珠法PCR产物纯化 片段化 Fragmentation Fragmented PCR Products Run on 4% TBE Agarose Gel at 5 V/cm for 45 minutes. Fragmentation is confirmed by majority of distribution between 25 to 125 bp 标记 Labeling 杂交 Hyb 上样 上样 GeneChip? Hybridization Oven 645 SNP array的技术原理 微阵列 SNP芯片含有大量寡核苷酸探针，起初设计被用于检测SNP等位基因。 SNP芯片不仅能够研究拷贝数变异（CNV），而且还能够对SNP基因型进行分析。 几个概念 Long contiguous stretches of homozygosity(LCSHs) 长片段连续绵延的纯合子 ranging in the size from 2.94 to 26.27 Mbp 遗传一致性区域(Identical by descent,IBD) 杂合性缺失(Loss of heterozygosity，LOH or Absence of heterozygosity,AOH) 单亲二倍体（Uniparental disomy） CytoScan? HD CytoScan? 750K CytoScan? Optima CytoScan HD Array CNV Probe: 195万 SNP Probe: 75万 共270万个探针 CytoScan? HD Array offers: Best coverage – The broadest coverage of constitutional, cancer, OMIM, and RefSeq genes on a single array for the most comprehensive and relevant cytogenetic analysis Highest performance – Exceeds cytogenetics community guidelines for specificity, sensitivity, and resolution across the genome Most SNPs – Confident breakpoint determination, allelic confirmation of copy number changes, high-resolution LOH, gene-level homozygosity mapping, and enhanced detection of low-level mosaics, sample heterogeneity, and triploidy CytoScan 750K Array CNV Probe: 55万 SNP Probe: 20万 共