Ultrasensitive determination of absolute mRNA amounts at attomole levels of nearly identical plant genes with high-throughput mass spectrometry (MassARRAY).
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.
Australian Genome Research Facility, Level 5, Gehrmann Laboratories, University of Queensland, St Lucia, Brisbane, Australia.
Rust
R
Turakulov
Sureeporn
S
Nontachaiyapoom
Keith R
KR
Mitchelson
Peter M
PM
Gresshoff
Artem E
AE
Men
D020869:Gene Expression Profiling / Q000379:methods*; D017343:Genes, Plant; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods; D012333:RNA, Messenger / Q000032:analysis*; D018749:RNA, Plant / Q000032:analysis; D012680:Sensitivity and Specificity; D013025:Glycine max / Q000235:genetics*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
D012333:RNA, Messenger; D018749:RNA, Plant