We have characterized the global gene expression patterns of arabidopsis pollen using serial analysis of gene expression sage. Serial analysis of gene expression sage is a powerful tool, which provides quantitative and comprehensive expression profile of genes in a given cell population. The principle is to isolate a unique sequence tags 910 bp in length from individual mrnas and linkage of tags serially into long dna molecules for lumpsum sequencing. Serial analysis of gene expression sage by sequencing. Sage allows you to digitally analyze gene expression patterns. Absolute quantitation techniques such as serial analysis of gene expression sage use tags, calculating the number of tags for a given sequence by the total number of tags. An evaluation of serial analysis of gene expression sage in the. Serial analysis of gene expression an overview sciencedirect. Serial analysis of gene expression sage is a powerful and efficient method for compiling quantitative. Zuelke1,3 biotechnology and germplasm laboratory3 and bovine functional genomics laboratory,4 animal and natural.
Serial analysis of gene expression nature protocols. We identified sets of genes expressed only or most abundantly in a specific stage of breast tumorigenesis. To comprehensively identify transcription start sites and the frequencies of individual mrnas in human cell libraries, a method of 5. Here we describe the serial analysis of gene expression sage, a technique that allowsa rapid, detailed analysis ofthousands of transcripts. Laske, samuel myllykangas, kazuo oshima, stefan heller. Serial analysis of gene expression sage is becoming a widely used gene expression profiling method for the study of development, cancer and other human diseases. To identify molecular alterations involved in the initiation and progression of breast carcinomas, we analyzed the global gene expression profiles of normal mammary epithelial cells and in situ, invasive, and metastatic breast carcinomas using serial analysis of gene expression sage. Request pdf serial analysis of gene expression serial analysis of gene expression sage is recently developed, a sequencedbased technique, which permits comprehensive and quantitative gene. Sage provides quantitative and comprehensive expression profiling in a given cell population. Serial analysis of gene expression identifies connective tissue growth factor expression as a prognostic biomarker in gallbladder cancer hector alvarez, alejandro corvalan, juan c. Serial analysis of gene expression predicts structural. The mrna is extracted from the organism, and reverse transcriptase is used to copy the mrna into.
A method was developed, called serial analysis of gene expression sage, that allows the quantitative and. Serial analysis of gene expression sage serial analysis of gene expression sage is an approach that allows rapid and detailed analysis of overall gene expression patterns. Another technology, serial analysis of gene expression sage, in part overcomes this limitation. Sage technique works by isolating short fragments of genetic information from the expressed genes that are present in the cell under study. Serial analysis of gene expression in adenocarcinoma. Rtpcr analysis 3, it evaluates only a limited number of genes at a time. Use of serial analysis of gene expression sage technology mikio yamamoto, toru wakatsuki, akiyuki hada, akihide ryo 1 department of biochemistry,national defense medical college,32namiki,tokorozawa,saitama 35985,japan abstract serial analysis of gene expression, or sage, is an experimental technique designed to gain a direct and.
Within the organisms, genes are transcribed and spliced in eukaryotes to produce mature mrna transcripts red. Not just of a few genes but for a cells complete gene expression profile. Sage starts with mrna and end with neat graphs that allow you to compare the gene expression of normal, developmental and diseased cells. I bet you can imagine plenty of times this would be useful. Using the serial analysis of gene expression technique, we surveyed transcriptomes of three major tissues panicles, leaves, and roots of a superhybrid rice oryza sativa strain, lyp9, in comparison to its parental cultivars, 9311 indica and pa64s japonica. To demonstrate this strategy, short diagnostic sequence tags were isolated from pancreas, concatenated, and cloned. A video by genome british columbia demonstrating gene expression. Serial analysis of gene expression sage is used to generate library of short sequence tags, each of which is then used to uniquely identify a transcript, with multiple tags forming long serial contigs across transcripts. Unlike many other techniques, sage allows both qualitative and quantitative analysis of previously unknown transcripts. Gene expression analysis typically involves the isolation or capture of transcribed rna within a sample, followed by amplification and subsequent detection and quantitation. It works by isolating short fragments of genetic information from the expressed genes that are present in the cell being studied. The steps involved in data analysis in serial analysis of gene. A sage serial analysis of gene expression view of breast.
Serial analysis of gene expression in the chicken otocyst journal of the association for research in otolaryngology, dec 2011 saku t. Serial analysis of gene expression sage is a technique that involves. Serial analysis of gene expression circulation research. Sage is an innovative technique that offers the potential.
The characteristics of an organism are determined by the genes expressed within it. Serial analysis of gene expression sage was used to generate a hippocampal expression profile of almost 30 000 genes in lal and sal mice. This molecular genetics lecture explains about gene expression analysis techniques using dna chip technology and microarrays. Serial transcriptomics experiments investigate the dynamics of gene expression changes associated with a quantitative variable such as time or dosage. We acquired 465,679 tags from the serial analysis of gene expression libraries, which were consolidated into 68,483 unique. Serial analysis of gene expression sage is an approach that allows rapid and detailed analysis of overall gene expression patterns. Pdf serial analysis of gene expression sage is a powerful. Unlike the alternative technologies of differential display and subtractive hybridization, sage produces a full catalog of all transcripts, not only differentially.
Serial analysis of gene expression sage internet archive. Using sage, we found differential expression of 191 genes. Sage produces a large library of short sequence tags that originate from mrna extracted from a tissue or purified cell sample. Serial analysis of gene expression sage in the skeletal muscle of pig. Serial analysis of gene expression sage is a patented, largescale mrnaprofiling technology that produces comprehensive, quantitative, and reproducible gene expression profiles originally described in refs. She also writes and publishes fiction, and in her free time enjoys yoga, biking, and taking care of her pets. Serial analysis of gene expression sage newsmedical. Serial analysis of gene expression sage ndeveloped in 1995 velculescu et al. Among those, serial analysis of gene expression sage stands out. Long serial analysis of gene expression for gene discovery and transcriptome profiling in the widespread marine coccolithophore emiliania huxleyi.
A method was developed, called serial analysis of gene expression sage, that allows the quantitative and simultaneous analysis of a large number of transcripts. First, a short nucleotide sequence tag 9 to 10 base pairs bp contains sufficient information. Serial analysis of gene expression in the chicken otocyst. The frequency of each tag directly reflects transcript abundance. Serial analysis of gene expression in turkey sperm storage. Serial analysis of gene expression in turkey sperm storage tubules in the presence and absence of resident sperm ezhou l. Pdf serial analysis of gene expression researchgate. Among these were genes involved in growth, signal transduction, and cell metabolism. The arabidopsis root transcriptome by serial analysis of. Serial analysis of gene expression sage is a common and powerful approach used for constructing expression libraries velculescu et al. Sage method allows for a quantitative and simultaneous analysis of a large number transcripts in any particular cells or tissues, without prior knowledge of the genes. Haemonchus contortus, serial analysis of gene expression, sage. Correction of technologyrelated artifacts in serial analysis of gene expression viatcheslav r. In the present study, genes with decreased expression in gc were screened for by serial analysis of gene expression sage data analysis and reverse transcription rt.
In the subsequent reaction, two different adaptors are ligated to free nlaiiidigested ends. Pdf the characteristics of an organism are determined by the genes expressed within it. A total of 21,237 sage tags were sequenced and 4,211 unique tags were identified. Over the course of the last years, sage has became a recognized tool of largescale gene expression profiling, being used extensively in human, animal. Serial analysis of gene expression in the silkworm. The serial analysis of gene expression sage method is based on the isolation of unique sequence tags from individual transcripts and concatenation of tags serially into long dna molecules. Total rna was resuspended in rnasefree water and evaluated by gel electrophoresis. Akmaev duplicate ditag analysis in longsage jeppe emmersen statistical comparison of two or.
Modified serial analysis of gene expression method for. Largescale identification of genes expressed in roots of the model plant arabidopsis was performed by serial analysis of gene expression sage, on a total of 144,083 sequenced tags, representing at least 15,964 different mrnas. Use of serial analysis of gene expression technology to. Serial analysis of gene expression, or sage, is an experimental technique designed to acquire direct quantification of gene expression. Serial analysis of gene expression identifies connective. The number of times each tag is detected is correlated to the expression level for the corresponding transcript. The major difference between microarray hybridization and serial analysis of gene expression sage techniques is that the latter does not require prior knowledge of the sequences to be analyzed, as sage is a sequencingbased gene expression profiling technique. Serial analysis of gene expression sage is a sequencebased approach that allows for global analysis of mrna transcripts, without the requirement of a priori knowledge of an organisms transcriptome. Serial analysis of gene expression study of a hybrid rice. Pdf serial analysis of gene expression in the silkworm. Serial analysis of gene expression sage was applied to the major cereal crop barley hordeum vulgare to characterize the transcriptional profile of grain during the malting process. Applications in malaria parasite, yeast, plant, and animal studies. Use of serial analysis of gene expression technology to reveal.