Subtractive hybridization is a technology that allows for
PCR-based amplification of only
cDNA
In genetics, complementary DNA (cDNA) is DNA synthesized from a single-stranded RNA (e.g., messenger RNA (mRNA) or microRNA (miRNA)) template in a reaction catalyzed by the enzyme reverse transcriptase. cDNA is often used to express a speci ...
fragments that differ between a control (driver) and experimental
transcriptome
The transcriptome is the set of all RNA transcripts, including coding and non-coding, in an individual or a population of cells. The term can also sometimes be used to refer to all RNAs, or just mRNA, depending on the particular experiment. The t ...
. cDNA is produced from
mRNA
In molecular biology, messenger ribonucleic acid (mRNA) is a single-stranded molecule of RNA that corresponds to the genetic sequence of a gene, and is read by a ribosome in the process of Protein biosynthesis, synthesizing a protein.
mRNA is ...
. 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
gDNA
Genomic deoxyribonucleic acid (abbreviated as gDNA) is chromosomal DNA, in contrast to extra-chromosomal DNAs like plasmids. Most organisms have the same genomic DNA in every cell; however, only certain genes are active in each cell to allow for c ...
s 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
Metagenomics is the study of genetic material recovered directly from environmental or clinical samples by a method called sequencing. The broad field may also be referred to as environmental genomics, ecogenomics, community genomics or microb ...
).
See also
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Representational difference analysis
Representational difference analysis (RDA) is a technique used in biological research to find sequence differences in two genomic or cDNA samples. Genomes or cDNA sequences from two samples (i.e. cancer sample and a normal sample) are PCR amplifi ...
External links
Overview at evrogen.com*
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Biotechnology
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