Single-cell laser-capture microdissection and RNA amplification

Methods Mol Med. 2004:99:215-23. doi: 10.1385/1-59259-770-X:215.

Abstract

Generating gene-expression profiles from laser-captured cells requires the successful combination of laser-capture microdissection, RNA extraction, RNA amplification, and microarray analysis. To permit single-cell gene-expression profiling, the RNA amplification method has to be sufficiently powerful to bridge the gap between the amount of RNA available from a single cell to what is required by the microarray, a gap that spans 5 to 6 orders of magnitude. This chapter focuses on the amplification of RNA using a two-round T7 RNA amplification method. The protocols described are adapted for laser-captured material and have been used to generate gene expression profiles from single laser-captured cells.

MeSH terms

  • Escherichia coli / genetics
  • Gene Expression Profiling / methods
  • Lasers*
  • Microdissection / instrumentation
  • Microdissection / methods*
  • Nucleic Acid Amplification Techniques / instrumentation
  • Nucleic Acid Amplification Techniques / methods*
  • Oligonucleotide Array Sequence Analysis / methods
  • RNA / genetics*

Substances

  • RNA