MicroSAGE: a modified procedure for serial analysis of gene expression in limited amounts of tissue

NA Datson*, J van der Perk-de Jong, MP van den Berg, ER de Kloet, E Vreugdenhil

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

132 Citations (Scopus)

Abstract

Serial Analysis of Gene Expression (SAGE) is a powerful expression profiling method, allowing the analysis of the expression of thousands of transcripts simultaneously, A disadvantage of the method, however, is the relatively high amount of input RNA required. Consequently, SAGE cannot be used for the generation of expression profiles when RNA is limited, i.e. in small biological samples such as tissue biopsies or microdissected material. Here we describe a modification of SAGE, named microSAGE, which requires 500- to 5000-fold less starting material. Compared with SAGE, microSAGE is simplified due to incorporation of a 'single-tube' procedure for all steps from RNA isolation to tag release, Furthermore, a limited number of additional PCR cycles are performed. Using microSAGE gene expression profiles can be obtained from minute quantities of tissue such as a single hippocampal punch from a rat brain slice of 325 mu m thickness, estimated to contain, at most, 10(5) cells. This method opens up a multitude of new possibilities for the application of SAGE, for example the characterization of expression profiles in tissue biopsies, tumor metastases or in other cases where tissue is scarce and the generation of region-specific expression profiles of complex heterogeneous tissues.

Original languageEnglish
Pages (from-to)1300-1307
Number of pages8
JournalNucleic Acids Research
Volume27
Issue number5
Publication statusPublished - 1-Mar-1999

Keywords

  • SUBTRACTIVE CDNA CLONING
  • ARBITRARILY PRIMED PCR
  • DIFFERENTIAL DISPLAY
  • MESSENGER-RNAS
  • SEQUENCE TAGS
  • HUMAN BRAIN
  • RAT
  • PROFILES
  • CELLS
  • IDENTIFICATION

Cite this