PPT-Restriction Enzyme digestion of DNA - Exercise 8

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. Richard J. Roberts. New England Biolabs. CSHL - . 2013. AHA!. Danna, K., Nathans, D. (1971) Specific cleavage of simian virus 40 DNA by restriction endonuclease of . Hemophilus. influenza. . Proc. Natl. Acad. Sci. USA 68: 2913-2917. . Restriction Enzymes. General Genetics. Objectives:. Introduce the students to digest genomic DNA by restriction . endonucleases. .. Observe the results of digestion on . agarose. gel electrophoresis.. An Introduction to Restriction Enzymes & Gel Electrophoresis. Objectives. Understand the use of restriction enzymes as biotechnology tools. Become familiar with principles and techniques of . agarose. Protocol 10.1 through 10.3. 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. Dr. Kevin Ahern. Protease Mechanisms. Aspartic Acid. Aspartic Acid. Histidine. Cysteine. Electron rich. Metal. Other Protease Families. Cysteine. Aspartyl. Metallo-. Papain. Cathepsin D. Carboxypeptidase A. are. . working. ?. Restriction. Enzymes. Natural . occurrence. . of. . restriction. . enzymes. . is. in . bacteria. . for. . protecting. . them. . against. a . virus. . attack. . by. . 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 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. . the isolation of the restriction . endonuclease. from. E. coli K (. Meselson. & Yuan 1968). . It was evident . that the restriction . endonucleases. from . E. coli K . and . E. coli B were important examples of proteins. Reaction: RE . DNA. Buffer. Suitable Temperature. All REs can be inactivated at high temperature . catalyzes the hydrolysis of the . phosphodiester. bond between adjacent nucleotides.. Choi Y, Jang W, Kim J, Ryu J, Lee S, Park K, et al. Spotted Fever Group and Typhus Group Rickettsioses in Humans, South Korea. Emerg Infect Dis. 2005;11(2):237-244. https://doi.org/10.3201/eid1102.040603. 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. Steps in RDT:. by. Prof. . Meena. . Nagawanshi. Professor . Department of . Zoology. Deogiri . College, Aurangabad. Enzymes. Enzymes are proteins. . biological catalysts .  help drive biochemical reactions. Enzyme names end with an .