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Serial analysis of gene expression:SAGE <br />
SAGE<br />SAGE is a powerful tool that allows the analysis of overall gene expression patterns with digital analysis.<br /...
SAGE PRINCIPLE<br />A short sequence tag (10-14bp) contains sufficient information to uniquely identify a transcript provi...
SAGE ADVANTAGE<br />Linking the tags together allows for rapid sequencing analysis of multiple transcripts.<br />By linkin...
SAGE METHODOLOGY<br /> Generate cDNA primed with biotin-oligo(dT)<br />NlaIII<br />Linker have restriction site for “taggi...
Schematic of SAGE method: <br />
What does the data look like?<br />
SAGE APPLICATION <br />SAGE is useful in comparative expression studies to identify differences in gene expression between...
SAGEmap<br />
http://www.sagenet.org/<br />
Sage resources<br />
Sage data<br />
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Sage

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Sage

  1. 1. Serial analysis of gene expression:SAGE <br />
  2. 2. SAGE<br />SAGE is a powerful tool that allows the analysis of overall gene expression patterns with digital analysis.<br /> Because SAGE does not require a preexisting clone, it can be used to identify and quantitate new genes as well as known genes. <br />
  3. 3. SAGE PRINCIPLE<br />A short sequence tag (10-14bp) contains sufficient information to uniquely identify a transcript provided that that the tag is obtained from a unique position within each transcript; <br />Sequence tags can be linked together to from long serial molecules that can be cloned and sequenced; and <br />Quantitation of the number of times a particular tag is observed provides the expression level of the corresponding transcript. <br />
  4. 4. SAGE ADVANTAGE<br />Linking the tags together allows for rapid sequencing analysis of multiple transcripts.<br />By linking the tags together, only one sequencing event is required to sequence every transcript within the cell.<br />Makes the task of DNA expression profiling a much easier<br />
  5. 5. SAGE METHODOLOGY<br /> Generate cDNA primed with biotin-oligo(dT)<br />NlaIII<br />Linker have restriction site for “tagging enzyme”(BsmFI)<br />
  6. 6.
  7. 7. Schematic of SAGE method: <br />
  8. 8. What does the data look like?<br />
  9. 9. SAGE APPLICATION <br />SAGE is useful in comparative expression studies to identify differences in gene expression between two or more cellular sources of RNA.<br /> A big advantage of the method is its speed: all genes expressed at 0.5% of total mRNA in one tissue but not abundant in another could be found in a single day<br />Approximately 1000 sequence tags from each tissue.<br />
  10. 10. SAGEmap<br />
  11. 11. http://www.sagenet.org/<br />
  12. 12. Sage resources<br />
  13. 13. Sage data<br />

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