Whither the Ideal RNA Amplification Kit?

Audrey Player is searching for the ideal RNA amplification protocol.

Written byJeff Minerd

| 6 min read

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This RNA-amplification protocol uses "terminal continuation" primers to enable production of either the standard antisense strand or the novel sense strand (shown). (From: S. Che, S.D. Ginsberg, Lab Invest., 84:131–7, 2004.)

Audrey Player is searching for the ideal RNA amplification protocol. In her microarray facility at the National Cancer Institute's Advanced Technology Center, she plans to genetically profile various subtypes of tumor cells obtained by laser-capture microdissection. The goal: to identify new targets for anticancer therapies.

But before she can begin, Player must find a protocol that can reliably amplify as little as 10 picograms of total input RNA from a single cell to the 5 or 20 micrograms necessary for microarray analysis (an increase of 107- to 108-fold) without distorting the original abundance ratios of each mRNA species. "Perhaps the most challenging aspect of studying the biology of small sample preparations or individual cells – what ...

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