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cDNA microarray analysis of small plant tissue samples using a cDNA tag target amplification protocol
Author(s) -
Hertzberg Magnus,
Sievertzon Maria,
Aspeborg Henrik,
Nilsson Peter,
Sandberg Göran,
Lundeberg Joakim
Publication year - 2001
Publication title -
the plant journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.058
H-Index - 269
eISSN - 1365-313X
pISSN - 0960-7412
DOI - 10.1046/j.1365-313x.2001.00972.x
Subject(s) - complementary dna , biology , microarray , phloem , dna microarray , gene expression profiling , rna , computational biology , gene expression , microarray analysis techniques , microbiology and biotechnology , gene , genetics , botany
Summary Microarray technology is becoming an important comprehensive tool to study gene expression in plants. However, the use of this technology is limited by the large amount of sample tissue needed for microarray analysis. Generally, 50–200 µg of total RNA and 1–2 µg of mRNA is required for each hybridisation, which is equivalent to 50–100 mg of plant tissue. This requirement for large amounts of starting material severely constrains the use of microarrays for transcript profiling in specific tissues and cell types during plant development. Here we report on a robust and reliable target amplification method that enables transcript profiling from sub‐mg amounts of plant tissue. Using 0.1 µg of total RNA we show that twofold expression differences are possible to distinguish with 99% confidence. We also demonstrate the application of this method in an analysis of secondary phloem development in hybrid aspen using defined tissue sections, corresponding to 2–4 cell layers with a fresh weight of ∼0.5 mg.