Suitability of Stratagene reference RNA for analysis of lymphoid tissues

• Published in: BioTechniques Vol. 37; no. 3; pp. 470 - 474
• Main Authors: Dybkær, Karen, Zhou, Guimei, Iqbal, Javeed, Kelly, David, Xiao, Li, Sherman, Simon, d'Amore, Francesco, Chan, Wing C
• Format: Journal Article
• Language: English
• Published: Natick, MA Future Science Ltd 01.09.2004; Eaton; Taylor & Francis Group
• Subjects: Biological and medical sciences; Carbocyanines; Cell Line, Tumor; Diverse techniques; Fluorescent Dyes; Fundamental and applied biological sciences. Psychology; Gene Expression Profiling - methods; Humans; Lymphoid Tissue; Molecular and cellular biology; Oligonucleotide Array Sequence Analysis; Oligonucleotide Probes; Reference Standards; Reproducibility of Results; RNA - genetics; RNA - isolation & purification; RNA - standards; Tissue; RNA; Analysis; Reference
• ISSN: 0736-6205; 1940-9818
• DOI: 10.2144/04373RT03

We evaluated a lymphoid RNA standard prepared in our laboratory for spotted microarrays against the Universal Human Reference standard from Stratagene. Our goal was to determine if the Stratagene standard, which contains only two lymphoid cell lines out of a pool of 10 human cancer cell lines, had acceptable gene coverage to serve as a comprehensive standard for gene expression profiling of lymphoid tissues. Our lymphoid standard was prepared from thymus, spleen, tonsil, and cell lines representing immature B cells, plasma cells, and natural killer (NK) cells, thus covering the entire spectrum of lymphoid cells and most stromal elements present in specialized lymphoid tissues. The two standards were co-hybridized on oligonucleotide microarrays containing 17,260 genes, and both had fluorescence intensities above background for approximately 85% of the genes. Despite the limited representation of lymphoid cells in the Stratagene standard, only 4.2% genes exhibited expression differences greater than 2-fold including only 0.35% with differences greater than 4-fold. Although the lymphoid standard reflected a more comprehensive representation of immune system-associated genes, the Stratagene standard has the advantage of being commercially available, enabling easier comparison across laboratories and allowing comparative studies across a long period of time.