PPT-Chapter 6:Plasmids 1 http://www.addgene.org/mol-bio-reference/plasmid-background/

Part 4 How do we isolate the plasmid DNA from our bacteria Isolating plasmid DNA The key is to separate the plasmids from the bacterial chromosome This is not trivial since they are both DNA What is the difference. PLASMID VECTORS. . . Cloning into a Plasmid. Bacteria are useful hosts.. They are easily grown. They are cheap to grow. They grow fast. They are easily manipulated in the laboratory. DNA can be inserted - transformation. It typically exists as a covalently closed circular piece of double stranded DNA that has the capability of replicating autonomously and it is this property that leads to its isolation and physical recognition. The closed covalent nature of their structure allows them to be separated from chromosomal DNA by either gel electrophoresis or . Objective. Students will model the process of using restriction enzymes and plasmids to form recombinant DNA.. Background Information. major tools of recombinant DNA technology are bacterial enzymes called restriction enzymes.. Dr. E. What is a plasmid?. Small double stranded circular DNA. Naturally found in bacteria. It is completely separate form bacterial chromosomal DNA. It independently self replicates . May give bacteria a . Figure 20.2. Bacterium. Bacterial. chromosome. Plasmid. 2. 1. 3. 4. Gene inserted into. plasmid. Cell containing gene. of interest. Recombinant. DNA (plasmid). Gene of . interest. Plasmid put into. bacterial cell. Lecture #9. Lehninger. . Principles of Biochemistry. Chapter 9. DNA-based information technologies. Genomics & Proteomics. Genome. : . A genome is the complete set of genetic information in an organism.. 118 Use the information provided below can be used to create a standard curve. From this curve you can and determine the size of the restriction fragments for the plasmid you are studying. Figure 1. A vehicle used to transfer genetic material such as DNA sequence from the donor organism to target cell of recipient organism(Host cell).. Also referred as carrier of DNA sequence.. Also referred as cloning vehicle or cloning DNA.. Jasmin . Sutkovic. Principles . of cloning, vectors and cloning strategies. by . Jasmin SUTJOVIC. Principles of cloning, vectors and cloning strategies. DNA CLONING. DNA cloning is a technique for reproducing DNA fragments. . Definition. Steps . Applications. INTRODUCTION. Recombinant DNA.  (. rDNA. ): .  . DNA.  molecules formed by laboratory methods of . genetic recombination.  (such as . molecular cloning. ) to bring together genetic material from multiple sources, creating . The primary Sigma product number covered by this technical bulletin is:CRISPR for: Custom gRNA expression plasmids (including All-in-One and gRNA-only plasmids) T7-generated gRNA Lentiviral particl S. mall . circular DNA molecules that replicate independently of the host chromosomes. How are plasmids constructed?. What functional elements are found in our yeast overexpression plasmids?. How are plasmids purified?. oriC. . gene, along with a . dnaA. . gene and other genes typical of the chromosomal origin of replication.. Plasmids usually carry genes for proteins that are necessary or beneficial to the host under some situations but are not essential under all conditions. . and phages . and are known as . phasmids. or, if they . contain . an . M13 . ori region. , phagemids. .. 1. . Phagemid. . vectors are plasmids having a small segment of a filamentous phage M-13, . fd.