Precious Cells Contain Precious Information

Strategies and Pitfalls in Expression Analysis from a Few Cells
  • Isabelle M. Henry
  • Dina F. Mandoli
Part of the Methods in Molecular Biology™ book series (MIMB, volume 236)

Abstract

Expression analysis, often encompassed in the term “functional genomics,” is the link between physiology and molecular biology. Often, specific physiological changes in plant development are due to a limited number of genes, expressed exclusively in very few cells of an organ or organism. Compounding the situation, these physiological changes may also be transient. Therefore, searching for the responsible genes, though exciting and necessary to understand important processes, is hindered primarily by the scarcity of “precious” cells in the desired physiological state. Used judiciously, molecular methods such as reverse transcription polymerase chain reaction (RT-PCR), microarray analysis, or subtractive hybridization allow analysis of rare or special cells. Each of these methods has its advantages and pitfalls. Working with precious cells entails special biological strategies to avoid excessive work in obtaining the data and misinterpretation of it. To illustrate the logic and methods involved in working with precious cells-tissues, we describe how subtractive hybridization followed by expressed sequence tag (EST) sequencing can be used to search for a few genes specific to a few available cells.

Key Words

EST Acetabularia developmental biology mRNA extraction suppressive subtractive hybridization bioinformatics normalization RT-PCR 

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Copyright information

© Humana Press Inc., Totowa, NJ 2003

Authors and Affiliations

  • Isabelle M. Henry
    • 1
  • Dina F. Mandoli
    • 1
    • 2
  1. 1.Department of BiologyUniversity of WashingtonSeattle
  2. 2.Center for Developmental BiologyUniversity of WashingtonSeattle

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