Comparison of oligonucleotide-microarray and serial analysis of gene expression (SAGE) in transcript profiling analysis of megakaryocytes derived from CD34+ cells

Exp Mol Med. 2003 Oct 31;35(5):460-6. doi: 10.1038/emm.2003.60.

Abstract

For the comprehensive analysis of transcript expression, the array-based hybridization analysis and the serial analysis of gene expression (SAGE) are commonly used platforms. The SAGE is based on a high-throughput sequencing of ditags derived from the transcript. DNA microarrays are a powerful tool for monitoring thousands of transcripts simultaneously, whereas the Genechip (Affimatrix microarray) technology is based on the hybridization of a single probe or other manufacturer's microarrays (cDNA- or oligonucleotide-microarray) procedures include the competitive hybridization of two probes. In this study, the quantitative accuracy of expression using oligonucleotide-microarray was determined by comparing data set from the SAGE. In previous study the microSAGE was performed for the megakaryocytes and non- megakaryocytes derived from human cord blood CD34+ cells by ex vivo expansion using thrombopoietin, and a total of 38,909 tags representing 8,976 unique genes were obtained. On the identical RNA, expression profiling was also carried out using oligonucleotide-microarray (MAGIC II 10K chip, Macrogen). The most frequently expressed genes in human megakaryocytes were identified as platelet factor 1 followed by annexin A1, ribosomal protein S23. The majority of the 50 most highly expressed genes in the CD34+-derived megakaryocytes were those involved in protein synthesis, e.g., ribosomal proteins. The expression level through the single channel of oligonucleotide-microarray and SAGE have a fairly good correlation in terms of absolute analyses and that the correlation is higher for the genes with higher expression levels.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antigens, CD34 / metabolism*
  • Fetal Blood
  • Gene Expression Profiling / methods*
  • Humans
  • Megakaryocytes / metabolism*
  • Oligonucleotide Array Sequence Analysis / methods*
  • RNA, Messenger / analysis
  • RNA, Messenger / genetics
  • Reproducibility of Results
  • Transcription, Genetic / genetics*

Substances

  • Antigens, CD34
  • RNA, Messenger