PPT-Chapter 6 Restriction enzyme cloning

Plasmid 1 Gene Plasmid 1 Gene Restriction Endonuclease Plasmid 2 Gene Restriction Endonuclease Ligation Gene A plasmid used for bluewhite selection 2 Full size 2961 bp Each cell contains only . 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. Gene Cloning . allows the separation and identification of a specific section of genetic . material . (DNA or RNA) from other sequences. . It then . allows the isolation of large numbers of copies . of . 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. Gene Cloning . allows the separation and identification of a specific section of genetic . material . (DNA or RNA) from other sequences. . It then . allows the isolation of large numbers of copies . of . Recombinant . DNA technology. Methods . used to join together (recombine) different DNA segments that are not found together in nature.. This technique is used in genetic analysis to serve several applications:. 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 A r t i c l e b y - , W e s t B e n g a l , I n d i a Emailed by Monazza Shahab A r t i c l e b y - M o n a z z a S h a h a b , W e s t B e n g a l , I n d i a CONSTRUCTION OF PARTIAL GENOMIC LIBRARY 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. 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 .