Suppression subtractive hybridization
Subtractive hybridization is a technology that allows for PCR-based amplification of only cDNA fragments that differ between a control (driver) and experimental transcriptome. cDNA is produced from mRNA. Differences in relative abundance of transcripts are highlighted, as are genetic differences between species. The technique relies on the removal of dsDNA formed by hybridization between a control and test sample, thus eliminating cDNAs or gDNAs of similar abundance, and retaining differentially expressed, or variable in sequence, transcripts or genomic sequences.
Suppression subtractive hybridization has also been successfully used to identify strain- or species-specific DNA sequences in a variety of bacteria including Vibrio species (Metagenomics).
See also
- Representational Difference Analysis
External links
- Overview at evrogen.com
- Munir, S.; Singh, S.; Kaur, K.; Kapur, V. (2004). "Suppression subtractive hybridization coupled with microarray analysis to examine differential expression of genes in virus infected cells". Biological Procedures Online 6: 94–104. doi:10.1251/bpo77. PMC 420231. PMID 15181476.
- Diatchenko, L. (1996). "Suppression subtractive hybridization: A method for generating differentially regulated or tissue-specific cDNA probes and libraries". Proceedings of the National Academy of Sciences 93 (12): 6025–6030. doi:10.1073/pnas.93.12.6025.
- Galbraith, E. A.; Antonopoulos, D. A.; White, B. A. (2004). "Suppressive subtractive hybridization as a tool for identifying genetic diversity in an environmental metagenome: The rumen as a model". Environmental Microbiology 6 (9): 928–937. doi:10.1111/j.1462-2920.2004.00575.x. PMID 15305918.