Expression profiling using a hexamer-based universal microarray
| Autor: | Stephen W Powell, Anthony Valerio, Kevin J. Mcconnell, Michelle Lacey, Li Feng, Paul M. Lizardi, Chris Michaelson, Barbara Jedrzkiewicz, Junhyong Kim, Brent A. Orr, Jane S Merkel, Matthew E. Roth, David M. Kranz, Jayashree Hariharan, Joseph C. Kaufman, Lorri Guccione, Jonathan Hnath, Anton Tevelev, Carol D Graham, Maura J Ford, Craig E Parman, Brenda Zolla, Jason P Affourtit, Darin R. Latimer, Cesar E. Guerra, Conrad Halling, Debasish Raha, Kevin R Piper, Paul J. Schaffer, Harish Krishnaswamy, Jennifer Lane, Rixin Wang, Martin J. Mattessich, Ji Liao, Gino Intrieri, Lisa Guccione, Clotilde Perbost, Baoge Ying |
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| Rok vydání: | 2004 |
| Předmět: |
DNA
Complementary Sequence analysis T-Lymphocytes Biomedical Engineering Bioengineering DNA Fragmentation Saccharomyces cerevisiae Computational biology Biology Applied Microbiology and Biotechnology Transcriptome Mice chemistry.chemical_compound Complementary DNA Gene expression Image Processing Computer-Assisted Animals Humans RNA Messenger Transgenes Muscle Skeletal Promoter Regions Genetic 3' Untranslated Regions Gene Oligonucleotide Array Sequence Analysis Models Genetic Reverse Transcriptase Polymerase Chain Reaction Gene Expression Profiling Muscles Galactose DNA Restriction Enzymes Sequence Analysis DNA Molecular biology Gene expression profiling Restriction enzyme chemistry Molecular Medicine Algorithms DNA Biotechnology |
| Zdroj: | Nature Biotechnology. 22:418-426 |
| ISSN: | 1546-1696 1087-0156 |
| DOI: | 10.1038/nbt948 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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