Differential display (also referred to as DDRT-PCR or DD-PCR) is a laboratory technique that allows a researcher to compare and identify changes in gene expression at the

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 ...

level between two or more eukaryotic cell samples. It was the most commonly used method to compare expression profiles of two eukaryotic cell samples in the 1990s. By 2000, differential display was superseded by

DNA microarray A DNA microarray (also commonly known as DNA chip or biochip) is a collection of microscopic DNA spots attached to a solid surface. Scientists use DNA microarrays to measure the expression levels of large numbers of genes simultaneously or to ...

approaches. In differential display, first all the RNA in each sample is reverse transcribed using a set of

3′ Directionality, in molecular biology and biochemistry, is the end-to-end chemical orientation of a single strand of nucleic acid. In a single strand of DNA or RNA, the chemical convention of naming carbon atoms in the nucleotide pentose-sugar-ri ...

"anchored primers" (having a short sequence of deoxy-thymidine nucleotides at the end) to create a cDNA library for each sample, followed by PCR amplification using arbitrary 3′ primers for cDNA strand amplification together with anchored 3′ primers for RNA strand amplification, identical to those used to create the library; about forty arbitrary primers is the optimal number to transcribe almost all of the mRNA. The resulting transcripts are then separated by

electrophoresis Electrophoresis, from Ancient Greek ἤλεκτρον (ḗlektron, "amber") and φόρησις (phórēsis, "the act of bearing"), is the motion of dispersed particles relative to a fluid under the influence of a spatially uniform electric fie ...

and visualized, so that they can be compared. The method was prone to error due to different mRNAs migrated into single bands, differences in less abundant mRNAs getting drowned by more abundant mRNAs, sensitivity to small changes in cell culture conditions, and a tendency to amplify 3′ fragments rather than full mRNAs, and the necessity to use about 300 primers to catch all the mRNA. The method was first published in ''Science'' in 1992.

References

{{DEFAULTSORT:Differential Display Biotechnology