Lecture 3.12 Notes

Lecture 3.12 Notes - Notes Genetic Engineering Tools and...

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Notes – 3/12/10 Genetic Engineering Tools and techniques Restriction and modification enzymes: Three major classes Widespread in prokaryotes – both Bacteria and Archaea o Rare in Eukaryotes Types I and III – bind at recognition sequences but cleave elsewhere Type II – bind and cleave at recognition sequences – much more useful for cloning Recognition of palindromes o Homodimeric proteins Size of recognition sequence frequency of cutting 4 n Modification enzymes: protection from restriction Gel electrophoresis – size separation of DNA molecules Ethidium bromide visualization of DNA Restriction endonuclease mapping Southern blot analysis Essentials of molecular cloning Isolation of specific genes in pure form Complexity of prokaryotic versus eukaryotic genomes Steps in gene cloning Isolation and fragmentation of source DNA Inserting DNA into appropriate cloning vector Introduction of cloned DNA into host organism Identification of desired clone development of specific probes Plasmids as cloning vectors
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