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RNArticles: RNA Amplification Used to Develop a New In Vitro HCMV Assay


Gene arrays can be used to simultaneously measure the expression levels of thousands of genes in a single experiment. One limitation of this technology is that large amounts of RNA may be required for each array hybridization experiment -- typically 10 g of total RNA. This becomes problematic when working with limited samples, such as small biopsies, mRNA-poor cells and tissues, primary cultured cells, and laser capture microdissection (LCM) samples, and, in this example, cells selected and collected by flow cytometry. RNA amplification is needed to perform array analysis on these limited samples.

Goodrum FD, Jordan CT, High K & Shenk T. (2002) Human cytomegalovirus gene expression during infection of primary hematopoietic progenitor Cells: A model for latency. Proc Natl Acad Sci (USA) 99 (25): 16255-16260.

Ambion's MessageAmp aRNA Amplification Kit is based on an antisense RNA (aRNA) amplification procedure first described by Van Gelder and Eberwine (1). The protocol involves a series of enzymatic reactions resulting in linear amplification of exceedingly small amounts of RNA for use in array analysis. Here this method is applied to amplify RNA from a small number of human cytomegalovirus (HCMV) infected CD34+ progenitor cells to compare the expression pattern of these infected cells, an in vitro model, with that of normal fibroblasts and to
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TAG: RNArticles RNA Amplification Used Develop New Vitro HCMV Assay

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