Serial analysis of gene expression
Serial analysis of gene expression
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DOI：
10.1038/nprot.2006.269
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发表时间：
2006-01-01
期刊：
影响因子：
14.8
通讯作者：
Polyak, Kornelia
中科院分区：
文献类型：
--
作者：
Hu, Min;Polyak, Kornelia
基因表达序列分析（SAGE）是一种获得全面、公正和定量的基因表达谱的方法。与阵列相比，它的主要优点是它不需要对待分析基因的先验知识，并且反映绝对的mRNA水平。由于最初的SAGE协议是以短标签（10-bp）格式开发的，因此已经进行了几次修改，以产生更长的SAGE标签，以便更精确地进行基因鉴定，并减少所需起始材料的数量。一些类似sage的方法也被开发用于DNA拷贝数变化和甲基化模式、染色质结构和转录因子靶点的全基因组分析。在本协议中，我们描述了针对少量起始材料优化的17-bp longSAGE转录组分析方法。这些文库的生成可以在7-10d内完成，而测序和数据分析则需要额外的2-3周。
Serial analysis of gene expression ( SAGE) is a method used to obtain comprehensive, unbiased and quantitative gene-expression profiles. Its major advantage over arrays is that it does not require a priori knowledge of the genes to be analyzed and reflects absolute mRNA levels. Since the original SAGE protocol was developed in a short-tag (10-bp) format, several modifications have been made to produce longer SAGE tags for more precise gene identification and to decrease the amount of starting material necessary. Several SAGE-like methods have also been developed for the genome-wide analysis of DNA copy-number changes and methylation patterns, chromatin structure and transcription factor targets. In this protocol, we describe the 17-bp longSAGE method for transcriptome profiling optimized for a small amount of starting material. The generation of such libraries can be completed in 7-10d, whereas sequencing and data analysis require an additional 2-3 wk.