Stages of Genetic Engineerin1

Stages of Genetic Engineerin1 - Stages of Genetic...

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Stages of Genetic Engineering DNA cleavage (stage 1) - restriction endonuclease cleaves DNA into fragments produces large number of different fragments different endonucleases >> different fragments gel electrophoresis - procedure that separates fragments based on size recombinant DNA production (stage 2) - DNA fragments inserted into vectors vectors cleaved w/ same restriction endonuclease as DNA cloning (stage 3) - more recombinant DNA created vectors introduced into reproducing cells each reproduced cell contains recombinant DNA screening (stage 4) - most challenging part of any genetics experiment 4-I - preliminary screening of clones o gets rid of cells w/o any vectors, vectors w/o original DNA o uses vector w/ gene for antibiotic resistance o based on presence/absence of a certain phenotype 4-II - finding gene of interest o hybridization - uses complementary nucleic acid ( probe ) to find particular fragment o solution added to denature DNA, allowing radioactive probe to attach
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