PPT-Restriction Enzyme Digestion of Phage DNA

Protocol 101 through 103 Objective To cut phage genome into multiple fragments based on DNA sequence General Introduction on Restriction Enzymes Are also known as restriction endonucleases Are naturally occurring enzymes used by bacteria for defensive purposes against extraneous DNA molecules. Bio-Rad Biotechnology . Explorer™ . DNA Fingerprinting Kit. Crime Scene. Have fun setting up your own crime scene. Be as elaborate or as simple as you wish.. Dye Electrophoresis . Could you eliminate any suspects using dye electrophoresis?. Restriction Enzymes. General Genetics. Objectives:. Introduce the students to digest genomic DNA by restriction . endonucleases. .. Observe the results of digestion on . agarose. gel electrophoresis.. Presented by Fotios Patsalis . Bio . 446/546 FALL 2016. M/F 13.00-15.50h,. Castetter. . Hall 37. Restriction Enzymes: Molecular Scissors. Restriction enzymes (. endonuleases. ) are specific endonucleases which . Objectives. -Understand how Restriction Enzymes digest DNA.. -Know how to construct a . pAMP. . (plasmid) or gel.. -Given the size of fragments, gel, know how to construct a restriction map.. -Given a restriction map know how to construct a gel.. Restriction enzymes. Polymerase enzyme. Strand-displacing polymerases. Helicase enzymes. Discovery and Obtainability:. Most enzymes are proteins discovered in cells.. But . DNAzymes. were . descovered. are. . working. ?. Restriction. Enzymes. Natural . occurrence. . of. . restriction. . enzymes. . is. in . bacteria. . for. . protecting. . them. . against. a . virus. . attack. . by. . Explorer™ . DNA Fingerprinting Kit. Crime Scene. Have fun setting up your own crime scene. Be as elaborate or as simple as you wish.. Dye Electrophoresis . Could you eliminate any suspects using dye electrophoresis?. Presented . by. Ms. . P. . . H. . Giri. Department of Microbiology. Deogiri . College, Aurangabad. B.Sc. T. Y. . Semester VI. Paper No. XIX. Recombinant DNA Technology. Ms. . Priyanka. H. . Giri. DNA manipulating enzymes:. Primer Design Molecular cloning requires manipulation of nucleic acid, including copying (synthesize), cutting (digest) and pasting (ligate). In bacteria, it is usually done in plasmid form. To inse M13. , f1 and . fd. are filamentous . coliphages. . containing a . circular . single-stranded DNA molecule. . These . coliphages. . have . been developed as cloning . vectors, for . they have a . number . Restriction enzymes . Restriction enzymes . (or restriction endonucleases), are bacterial enzymes that cleave both strands of DNA at or near specific recognition nucleotide sequences (4 -8 bp ) known as restriction sites. . Restriction Modification System. Cutting DNA molecules. . Before 1970s, there was no method of cleaving DNA at the specific points.. All the available methods for fragmenting DNA were non-specific.. or phages are viruses that specifically infect bacteria. . Viral vectors are those in which gene of interest is incorporated in the genome of virus.. The phage particle attaches to the outer surface of bacterium and injects its DNA into the cell.. Cutting DNA molecules. . Before 1970s, there was no method of cleaving DNA at the specific points.. All the available methods for fragmenting DNA were non-specific.. Mechanical shearing.