Quantitative cDNA-AFLP analysis for genome-wide expression studies
- Author
- Peter Breyne (UGent) , Rozemarijn Dreesen, Bernard Cannoot (UGent) , Debbie Rombaut (UGent) , Klaas Vandepoele (UGent) , Stephane Rombauts (UGent) , Rudy Vanderhaeghen (UGent) , Dirk Inzé (UGent) and Marc Zabeau (UGent)
- Organization
- Abstract
- An improved cDNA-AFLP method for genome-wide expression analysis has been developed. We demonstrate that this method is an efficient tool for quantitative transcript profiling and a valid alternative to microarrays. Unique transcript tags, generated from reverse-transcribed messenger RNA by restriction enzymes, were screened through a series of selective PCR amplifications. Based on in silico analysis, an enzyme combination was chosen that ensures that at least 60% of all the mRNAs were represented by an informative sequence tag. The sensitivity and specificity of the method allows one to detect poorly expressed genes and distinguish between homologous sequences. Accurate gene expression profiles were determined by quantitative analysis of band intensities, and subtle differences in transcriptional activity were revealed. A detailed screen for cell cycle-modulated genes in tobacco demonstrates the usefulness of the technology for genome-wide expression analysis.
- Keywords
- gene expression, microarrays, functional genomics, cell cycle, DISPLAY, CELL-CYCLE, transcriptome, HISTONE GENE-EXPRESSION
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Citation
Please use this url to cite or link to this publication: http://hdl.handle.net/1854/LU-211939
- MLA
- Breyne, Peter, et al. “Quantitative CDNA-AFLP Analysis for Genome-Wide Expression Studies.” MOLECULAR GENETICS AND GENOMICS, vol. 269, no. 2, 2003, pp. 173–79, doi:10.1007/s00438-003-0830-6.
- APA
- Breyne, P., Dreesen, R., Cannoot, B., Rombaut, D., Vandepoele, K., Rombauts, S., … Zabeau, M. (2003). Quantitative cDNA-AFLP analysis for genome-wide expression studies. MOLECULAR GENETICS AND GENOMICS, 269(2), 173–179. https://doi.org/10.1007/s00438-003-0830-6
- Chicago author-date
- Breyne, Peter, Rozemarijn Dreesen, Bernard Cannoot, Debbie Rombaut, Klaas Vandepoele, Stephane Rombauts, Rudy Vanderhaeghen, Dirk Inzé, and Marc Zabeau. 2003. “Quantitative CDNA-AFLP Analysis for Genome-Wide Expression Studies.” MOLECULAR GENETICS AND GENOMICS 269 (2): 173–79. https://doi.org/10.1007/s00438-003-0830-6.
- Chicago author-date (all authors)
- Breyne, Peter, Rozemarijn Dreesen, Bernard Cannoot, Debbie Rombaut, Klaas Vandepoele, Stephane Rombauts, Rudy Vanderhaeghen, Dirk Inzé, and Marc Zabeau. 2003. “Quantitative CDNA-AFLP Analysis for Genome-Wide Expression Studies.” MOLECULAR GENETICS AND GENOMICS 269 (2): 173–179. doi:10.1007/s00438-003-0830-6.
- Vancouver
- 1.Breyne P, Dreesen R, Cannoot B, Rombaut D, Vandepoele K, Rombauts S, et al. Quantitative cDNA-AFLP analysis for genome-wide expression studies. MOLECULAR GENETICS AND GENOMICS. 2003;269(2):173–9.
- IEEE
- [1]P. Breyne et al., “Quantitative cDNA-AFLP analysis for genome-wide expression studies,” MOLECULAR GENETICS AND GENOMICS, vol. 269, no. 2, pp. 173–179, 2003.
@article{211939,
abstract = {{An improved cDNA-AFLP method for genome-wide expression analysis has been developed. We demonstrate that this method is an efficient tool for quantitative transcript profiling and a valid alternative to microarrays. Unique transcript tags, generated from reverse-transcribed messenger RNA by restriction enzymes, were screened through a series of selective PCR amplifications. Based on in silico analysis, an enzyme combination was chosen that ensures that at least 60% of all the mRNAs were represented by an informative sequence tag. The sensitivity and specificity of the method allows one to detect poorly expressed genes and distinguish between homologous sequences. Accurate gene expression profiles were determined by quantitative analysis of band intensities, and subtle differences in transcriptional activity were revealed. A detailed screen for cell cycle-modulated genes in tobacco demonstrates the usefulness of the technology for genome-wide expression analysis.}},
author = {{Breyne, Peter and Dreesen, Rozemarijn and Cannoot, Bernard and Rombaut, Debbie and Vandepoele, Klaas and Rombauts, Stephane and Vanderhaeghen, Rudy and Inzé, Dirk and Zabeau, Marc}},
issn = {{1617-4615}},
journal = {{MOLECULAR GENETICS AND GENOMICS}},
keywords = {{gene expression,microarrays,functional genomics,cell cycle,DISPLAY,CELL-CYCLE,transcriptome,HISTONE GENE-EXPRESSION}},
language = {{eng}},
number = {{2}},
pages = {{173--179}},
title = {{Quantitative cDNA-AFLP analysis for genome-wide expression studies}},
url = {{http://doi.org/10.1007/s00438-003-0830-6}},
volume = {{269}},
year = {{2003}},
}
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