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. LOCK. it out on the first move. HI, DROXYLATE and . SWEET RELEASE defeat METHYLATE. but stay in play until cancelled. If host plays RESTRICT. then phage can counter with. METHYLATE – move both cards to the discard pile. 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. 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. 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.. 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. 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.. Reaction: RE . DNA. Buffer. Suitable Temperature. All REs can be inactivated at high temperature . catalyzes the hydrolysis of the . phosphodiester. bond between adjacent nucleotides.. 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.. 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:. 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.