Quantitation of gene copy number and mRNA using the polymerase chain reaction.

摘要: Quantitation of the copy number a specific gene in given sample on genomic as well steady state RNA transcriptional level is great interest both basic and clinical research. Traditionally, estimated by Southern blot or dot hybridization techniques. A certain amount total DNA digested with restriction enzymes, size fractionated nondenaturing agarose gel electrophoresis, transferred permanently bound to nitrocellulose nylon membrane (Southern blot); alternatively, can be without previous manipulations (dot blot). The then incubated radioactive, labeled molecular probe, which specifically binds interest. After washing steps remove unbound exposed film signal detected. intensity signal, quantitated (e.g., densitometry), correlates specific...