VB 550

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Credit: 4 hours.

Gives participants the background information and hands-on experience in the methodologies necessary to utilize cloned genes for the detection and quantitation of specific mRNA transcripts in RNA extracted from tissue or cell culture samples. Methodologies covered will include: recombinant plasmid propagation, cDNA probe isolation and isotopic labeling, RNA isolation, Poly A+ mRNA selection, gel separation and transfer of RNA to a membrane (Northern blot), hybridization of specific gene probes to membrane bound RNA (Northern hybridization), detection and quantitation of hybridization signal. These basic methodologies are widely applicable to different experimental systems. They allow an investigator to monitor the effects of physiological manipulations, to animals or cultured cells, at the molecular level. Prerequisite: Consent of instructor.

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