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Related Experiment Videos

Preparation of label from small cell numbers for microarray screening.

Heather L Wilson1, Helen C O'Neill

  • 1School of Biochemistry and Molecular Biology, Faculty of Science, Australian National University, Canberra, Australia.

Omics : a Journal of Integrative Biology
|January 27, 2005
PubMed
Summary

This study presents a method for amplifying labelled complementary RNA (cRNA) from minimal total RNA amounts. This enables gene expression profiling of rare cell populations using Affymetrix GeneChip technology.

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Area of Science:

  • Molecular Biology
  • Genomics
  • Biotechnology

Background:

  • Standard cRNA amplification for Affymetrix GeneChips requires 2-5 microg of total RNA, limiting studies on rare cells.
  • Previous workarounds like using whole tissue or cell lines have drawbacks, including cell contamination and lack of in vivo relevance.

Purpose of the Study:

  • To describe a method for amplifying labelled cRNA from low amounts of total RNA.
  • To enable gene expression analysis of rare cell subsets using Affymetrix GeneChip technology.

Main Methods:

  • Utilizing two cycles of amplification to generate labelled cRNA.
  • Starting with as little as 2 ng of total RNA.

Main Results:

  • Successful generation of sufficient labelled cRNA for hybridization from minimal starting material.

Related Experiment Videos

  • Enabling gene expression screening for low-number cells, rare cell subsets, and small patient biopsies.

    • Conclusions:

    • The described amplification method overcomes the limitation of high RNA input for Affymetrix GeneChip analysis.
    • This approach allows for accurate gene expression profiling of specific, rare cell populations of interest.