Journal Article

Ultrasensitive Determination of Absolute mRNA Amounts at Attomole Levels of Nearly Identical Plant Genes with High-Throughput Mass Spectrometry (MassARRAY)

Rust Turakulov, Sureeporn Nontachaiyapoom, Keith R. Mitchelson, Peter M. Gresshoff and Artem E. Men

in Plant and Cell Physiology

Published on behalf of Japanese Society of Plant Physiologists

Volume 48, issue 9, pages 1379-1384
Published in print September 2007 | ISSN: 0032-0781
Published online September 2007 | e-ISSN: 1471-9053 | DOI: http://dx.doi.org/10.1093/pcp/pcm103
Ultrasensitive Determination of Absolute mRNA Amounts at Attomole Levels of Nearly Identical Plant Genes with High-Throughput Mass Spectrometry (MassARRAY)

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  • Biochemistry
  • Molecular and Cell Biology
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Detection of very small amounts of RNA based on microdissection of plant tissue is essential for modern plant biology. Mass spectroscopy technology (MassARRAY) based on Sequenom™ instrumentation was adapted to determine quickly and in a high-throughput fashion (by multiplexing) the absolute amounts of mRNA of closely related soybean genes. A sensitivity of 0.1 amol (10−19) was achieved, representing as few as 1,000 mRNA molecules. This methodology eliminates the use of housekeeping genes as reference standards and has multiple applications for plant functional genomics, such as the monitoring of individual expression of paralogous genes at ultra-low expression levels and/or in extremely small tissue samples.

Keywords: Allele-specific expression — Competitive PCR — Gene expression — Multiplexing — MALDI TOF

Journal Article.  2667 words.  Illustrated.

Subjects: Biochemistry ; Molecular and Cell Biology ; Plant Sciences and Forestry

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