Expression profiling using a hexamer-based universal microarray.

Imagen de Paul Lizardi
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TítuloExpression profiling using a hexamer-based universal microarray.
Publication TypeJournal Article
Year of Publication2004
AutoresRoth, ME, Feng, L, McConnell, KJ, Schaffer, PJ, Guerra, CE, Affourtit, JP, Piper, KR, Guccione, L, Hariharan, J, Ford, MJ, Powell, SW, Krishnaswamy, H, Lane, J, Guccione, L, Intrieri, G, Merkel, JS, Perbost, C, Valerio, A, Zolla, B, Graham, CD, Hnath, J, Michaelson, C, Wang, R, Ying, B, Halling, C, Parman, CE, Raha, D, Orr, B, Jedrzkiewicz, B, Liao, J, Tevelev, A, Mattessich, MJ, Kranz, DM, Lacey, M, Kaufman, JC, Kim, J, Latimer, DR, Lizardi, PM
JournalNat Biotechnol
Volume22
Issue4
Pagination418-26
Date Published2004 Apr
ISSN1087-0156
Palabras clave3' Untranslated Regions, Algorithms, Animals, DNA Fragmentation, DNA Restriction Enzymes, DNA, Complementary, Galactose, Gene Expression Profiling, Humans, Image Processing, Computer-Assisted, Mice, Models, Genetic, Muscle, Skeletal, Muscles, Oligonucleotide Array Sequence Analysis, Promoter Regions, Genetic, Reverse Transcriptase Polymerase Chain Reaction, RNA, Messenger, Saccharomyces cerevisiae, Sequence Analysis, DNA, T-Lymphocytes, Transgenes
Abstract

We describe a transcriptional analysis platform consisting of a universal micro-array system (UMAS) combined with an enzymatic manipulation step that is capable of generating expression profiles from any organism without requiring a priori species-specific knowledge of transcript sequences. The transcriptome is converted to cDNA and processed with restriction endonucleases to generate low-complexity pools (approximately 80-120) of equal length DNA fragments. The resulting material is amplified and detected with the UMAS system, comprising all possible 4,096 (4(6)) DNA hexamers. Ligation to the arrays yields thousands of 14-mer sequence tags. The compendium of signals from all pools in the array-of-universal arrays comprises a full-transcriptome expression profile. The technology was validated by analysis of the galactose response of Saccharomyces cerevisiae, and the resulting profiles showed excellent agreement with the literature and real-time PCR assays. The technology was also used to demonstrate expression profiling from a hybrid organism in a proof-of-concept experiment where a T-cell receptor gene was expressed in yeast.

DOI10.1038/nbt948
Alternate JournalNat. Biotechnol.
PubMed ID15024387