Expression profiling using a hexamer-based universal microarray.
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Título | Expression profiling using a hexamer-based universal microarray. |
Publication Type | Journal Article |
Year of Publication | 2004 |
Autores | Roth, 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 |
Journal | Nat Biotechnol |
Volume | 22 |
Issue | 4 |
Pagination | 418-26 |
Date Published | 2004 Apr |
ISSN | 1087-0156 |
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. |
DOI | 10.1038/nbt948 |
Alternate Journal | Nat. Biotechnol. |
PubMed ID | 15024387 |