老化原因（英文版）.pptVIP

Vast amounts of data is produced, which must be sifted and ordered for useful information to become apparent. Sage reference databases: SAGE map SAGE Genie /cgap What does the data look like? FROM TAGS TO GENES…… Collect sequence records from GenBank Assign sequence orientation (by finding poly-A tail or poly-A signal or from annotations) Extract 10-bases -adjacent to 3’-most CATG Assign UniGene identifier to each sequence with a SAGE tag Record (for each tag-gene pair) #sequences with this tag #sequences in gene cluster with this tag Maps available at /SAGE DIFFERENTIAL GENE EXPRESSION BY SAGE Identification of differentially expressed genes in samples from different physiological or pathological conditions. Application of many statistical methods Poisson approximation Bayesian method Chi square test. SAGE software searches GenBank for matches to each tag This allows assignment to 3 categories of tags: mRNAs derived from known genes anonymous mRNAs, also known as expressed sequence tags (ESTs) mRNAs derived from currently unidentified genes SAGE VS MICROARRAY SAGE – An open system which detects both known and unknown transcripts and genes. COMPARISON…… SAGE Detects 3’ region of transcript. Restriction site is determining factor. Collects sequence information and copy no. Sequencing error and quantitation bias. MICROARRAY Targets various regions of the transcript.Base composition for specificity of hybridization. Fluorescent signals and signal intensity. Labeling bias and noise signals. Contd…… Features SAGE Microarray Detects unknown transcripts Yes No Quantification Absolute measure Relative measure Sensitivity High Moderate Specificity Moderate High Reproducibility Good for higher abundance transcripts Good for data from intra-platform comparison Direct cost 5-10X higher than arrays. 5-10 X lower than SAGE RECENT SAGE APPLICATIONS Analysis of yeast transcriptome Gene Expression Profiles in Normal and Cancer Cell Insights into p53-mediated apoptosis