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Related Experiment Videos

Serial analysis of gene expression.

Min Hu1, Kornelia Polyak

  • 1Department of Medical Oncology, Dana-Farber Cancer Institute, Harvard Medical School, 44 Binney Street, D740C, Boston, Massachusetts 02115, USA.

Nature Protocols
|May 10, 2007
PubMed
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Serial analysis of gene expression (SAGE) provides unbiased gene expression profiles without prior gene knowledge. This study details a modified 17-bp longSAGE method for efficient transcriptome profiling using minimal starting material.

Area of Science:

  • Molecular Biology
  • Genomics

Background:

  • Serial analysis of gene expression (SAGE) offers quantitative, unbiased gene expression profiling.
  • SAGE requires no prior gene knowledge and measures absolute mRNA levels, unlike array-based methods.
  • Modifications to SAGE have improved tag length for precise gene identification and reduced starting material requirements.

Purpose of the Study:

  • To describe an optimized 17-bp longSAGE protocol for transcriptome profiling.
  • To adapt SAGE for applications requiring minimal starting material.

Main Methods:

  • The study details the 17-bp longSAGE protocol.
  • This method is optimized for small amounts of starting material.
  • The protocol includes library generation, sequencing, and data analysis.

Related Experiment Videos

Main Results:

  • The 17-bp longSAGE method enables precise gene identification.
  • Library generation is efficient, completed in 7-10 days.
  • Sequencing and data analysis take an additional 2-3 weeks.

Conclusions:

  • The 17-bp longSAGE method provides a robust approach for transcriptome profiling.
  • This protocol is suitable for studies with limited starting material.
  • SAGE and its variants offer versatile tools for genomic analysis.