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#radioactive probes

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Gene Cloning Process

Which method uses radioactive probes to identify recombinant colonies?

Colony hybridization, developed by Grunstein and Hogness, identifies recombinant clones harboring specific DNA sequences without phenotypic screening. Colonies are replica-transferred to nitrocellulose or nylon membrane, lysed with alkali, and DNA denatured and fixed. The membrane is then incubated with a 32P or 35S labeled nucleic acid probe complementary to desired insert. After stringent washing to remove non-specific binding, positive colonies are detected by autoradiography where hybridization signal exposes X-ray film. Blue-white screening and direct antibiotic selection do not employ ra

Ref: NCERT Biology Class XII Principles on Klenow fill-in labeling, Lehninger Chapter 9 DNA cloning techniques, and Molecular Cloning by Sambrook Chapter 10 documenting end-labeling of cohesive termini.

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