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

Р. И. Туракулов | Zendy; Sureeporn tachaiyapoom | Zendy; Keith Mitchelson | Zendy; P.M. Gresshoff | Zendy; A. Men | Zendy

Open Access

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

Author(s) -

Р. И. Туракулов,

Sureeporn tachaiyapoom,

Keith Mitchelson,

P.M. Gresshoff,

A. Men

Publication year - 2007

Publication title -

plant and cell physiology

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 1.975

H-Index - 152

eISSN - 1471-9053

pISSN - 0032-0781

DOI - 10.1093/pcp/pcm103

Subject(s) - housekeeping gene , laser capture microdissection , gene , mass spectrometry , microdissection , throughput , biology , reference genes , microbiology and biotechnology , gene expression , computational biology , chemistry , genetics , chromatography , computer science , telecommunications , wireless

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 Sequenomtrade mark 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.

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