ABSTRACT
The in-gel DNA renaturation technique is the only method that allows one to detect the presence of amplified DNA of unknown nature by screening multiple DNA samples. Frequent occurrence of gene amplification in mammalian cell lines selected for various phenotypes, as well as in tumor cells, provides a unique opportunity for identification and cloning of unknown genes responsible for important cellular properties. The level of sensitivity achieved by SINE hybridization after removal of short duplexes is probably close to the absolute limit for any technique that detects amplified DNA sequences on the basis of their preferential reassociation. The cloned region of amplified DNA can then be tested for the presence of transcriptionally active sequences, and the identified transcribed fragments can be used as probes for cloning of the corresponding full-length cDNA sequences. In addition to detection and cloning of amplified DNA, the in-gel DNA renaturation technique can be used to analyze the structure of a known amplicon.
