Unit

UNIT 25B.9 Quantification of Gene Transcripts with Deep Sequencing Analysis of Gene Expression (DSAGE) Using 1 to 2 µg Total RNA

  1. Danos C. Christodoulou,
  2. Joshua M. Gorham,
  3. Masataka Kawana,
  4. Steven R. DePalma,
  5. Daniel S. Herman,
  6. Hiroko Wakimoto

Published Online: 1 JAN 2011

DOI: 10.1002/0471142727.mb25b09s93

Current Protocols in Molecular Biology

Current Protocols in Molecular Biology

How to Cite

C. Christodoulou, D., Gorham, J. M., Kawana, M., DePalma, S. R., Herman, D. S. and Wakimoto, H. 2011. Quantification of Gene Transcripts with Deep Sequencing Analysis of Gene Expression (DSAGE) Using 1 to 2 µg Total RNA. Current Protocols in Molecular Biology. 93:B:25B.9:25B.9.1–25B.9.16.

Author Information

  1. Harvard Medical School, Boston, Massachusetts

Publication History

  1. Published Online: 1 JAN 2011
  2. Published Print: JAN 2011

Abstract

Deep sequencing analysis of gene expression (DSAGE) measures global gene transcript levels from only 1 to 2 µg total RNA by massively parallel sequencing of cDNA tags. This unit describes the construction of 21-bp cDNA tag libraries appropriate for massively parallel sequencing and analysis of the resulting sequence data. The adapter oligonucleotides used are optimized for sequencing with current Illumina massively parallel sequencers, and a step-by-step implementation of the analysis protocol is described. The expression profiles obtained are highly reproducible, enabling sensitive detection of differences between experimental conditions as well as assessment of the relative transcript abundance of different genes. Curr. Protoc. Mol. Biol. 93:25B.9.1-25B.9.16. © 2011 by John Wiley & Sons, Inc.

Keywords:

  • digital gene expression;
  • serial analysis of gene expression (SAGE);
  • gene expression profiling;
  • high-throughput sequencing