Serial analysis of gene expression

Serial Analysis Of Gene Expression

Serial analysis of gene expression

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Description:
Serial analysis of gene expression (SAGE) is a technique used by molecular biologists to produce a snapshot of the messenger RNA population in a sample of interest in the form of small tags that correspond to fragments of those transcripts. The original technique was developed by Dr. Victor Velculescu at the Oncology Center of Johns Hopkins University and published in 1995. RL-SAGE and the most recent SuperSAGE. Many of these have improved the technique with the capture of longer tags, enabling more confident identification of a source gene.

Overview

Briefly, SAGE experiments proceed as follows:

  1. Isolate the mRNA of an input sample (e.g. a tumour).
  2. Extract a small chunk of sequence from a defined position of each mRNA molecule.
  3. Link these small pieces of sequence together to form a long chain (or concatemer).
  4. Clone these chains into a vector which can be taken up by bacteria.
  5. Sequence these chains using modern high-throughput DNA sequencers.
  6. Process this data with a computer to count the small sequence tags.


A more in-depth, technical explanation of the technique is available .

Analysis

The output of SAGE is a list of short sequence tags and the number of times it is observed. Using sequence databases a researcher can usually...
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