cDNA Subtraction

A difference cloning method and a technique for comparison of two closely related cell types, e.g. a differentiated or transformed cell (tester) and the undifferentiated or normal cell (driver). Driver RNA is converted to double-strand cDNA, usually with the incorporation of a tagged nucleotide, while the tester RNA is converted to first-strand cDNA (with no tag). The two preparations are mixed, melted and annealed to form, among other species, heteroduplexes composed of one strand from each of the different sources. The annealed heteroduplexes, as well as other driver species are captured by the tagged nucleotide and removed from the mixture. The resulting pool will be enriched for those cDNA species unique to the tester cells. Following second-strand synthesis, the double-stranded cDNAs are cloned. The resulting cDNA library will be enriched for sequences expressed only in the tester cells. cDNA subtraction can also be done by PCR. (NCI Thesaurus/OSP)

YOU MAY ALSO LIKE

cDNA Microarray Analysis

Complementary DNA (cDNA) clone inserts robotically printed onto a glass slide and subsequently hybridized to two differentially fluorescently labeled probes. The probes…

cDNA Library Construction

Methods for building collections of cloned DNA consisting of DNA copies of all the mRNAs produced by a cell type. cDNAs…

cDNA Library

A collection of DNA molecules that have been cloned in vectors. In the case of a cDNA library the DNA inserts…

cDNA Expression

CDNA expressed as protein formation

cDNA Array

A molecular biology device that utilizes a set of defined cDNA clones attached in a specific grid arrangement to a solid support…

cDNA

Single-stranded DNA that is complementary to messenger RNA or DNA that has been synthesized from messenger RNA by reverse transcriptase.