Identification of low-abundance differentially expressed transcripts using arrayed cDNA clones

• Published in: Comparative Biochemistry and Physiology Part B: Biochemistry and Molecular Biology Vol. 133; no. 4; pp. 537 - 542
• Main Authors: Golby, P, Stephens, S.K, Rast, J.P, Burke, J.F
• Format: Journal Article
• Language: English
• Published: England Elsevier Inc 01.12.2002
• Subjects: Animals; DNA array; DNA, Complementary; Drosophila melanogaster - genetics; Drosophila melanogaster - metabolism; Drosophila Proteins - genetics; Drosophila Proteins - metabolism; Female; Filter; Gene discovery; Gene expression; Gene Expression Profiling; Gene Library; Macroarray; Male; Membrane; Nucleic Acid Hybridization; Nylons; Oligonucleotide Array Sequence Analysis - methods; Polymerase Chain Reaction; RNA, Messenger - genetics; RNA, Messenger - metabolism; Subtractive hybridisation; Transcription, Genetic; Subtractive hybridisation; DNA array; Filter; Membrane; Macroarray; Gene expression; Gene discovery
• ISSN: 1096-4959; 1879-1107
• DOI: 10.1016/S1096-4959(02)00146-X

Analysis of comparative gene expression by the use of DNA microarrays has become a widely used tool. However, this technique is only readily applied to organisms where sequence information is known. This paper describes the development of a low-cost method of gene discovery by enrichment of differentially expressed transcripts, which uses cDNA library arrays of bacterial clones on nylon membranes (macroarrays) coupled with a subtractive probe preparation method to discover differentially expressed genes. The method requires no prior knowledge of the organism's genome sequence and overcomes the inherent insensitivity of standard methods of macroarray hybridisation.