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Plasmid Cloning Vectors Plasmids are circular, double-stranded DNA molecules that exist in bacteria and in the nuclei of some eukaryotic cells. They can replicate independently of the host cell.
Date added: July 18, 2013 - Views: 27
Cloning into Plasmids Restriction Fragment Cloning & PCR Cloning by the Topo TA™ Method Cloning Vectors The molecular analysis of DNA has been made possible by the cloning of DNA.
Date added: December 2, 2011 - Views: 81
Plasmids and Vectors Instructor Supplement to pGlo Bacterial Transformation A more detailed look at plasmids Cloning into a Plasmid Asilomar Conference Screening bacteria by replica plating Cosmids Other Vectors BACs (Bacterial artificial chromosomes) Large low copy number plasmids (have ori and ...
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Why Plasmids are Good Cloning Vectors small size (easy to manipulate and isolate) circular (more stable) replication independent of host cell several copies may be present (facilitates replication) frequently have antibody resistance ...
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Section 20.1. While . E. coli. is used as a prokaryotic host cell when working with plasmids, yeast is widely used as a host for DNA cloning and expression of eukaryotic genes because
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Plasmids and Cloning DNA By: Trevor Wade, Tori Marko and Melissa Dehghan What is DNA Cloning? A fragment of DNA that has been introduced into a foreign cell This results in exact copies of the original DNA fragment being made when the foreign cell replicates and divides Gene cloning became ...
Date added: July 15, 2013 - Views: 14
Plasmid Vectors Plasmids are circular pieces of DNA found naturally in bacteria. Plasmids can carry antibiotic resistance ... So, most cloning plasmids will contain two markers: An antibiotic marker that will allow only those bacteria with plasmids to grow on medium containing the appropriate ...
Date added: September 19, 2012 - Views: 66
Introduction to Vectors In order to study a DNA fragment (e.g., a gene), it needs to be amplified and eventually purified. These tasks are accomplished by cloning the DNA into a vector.
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Introducing our recombinant plasmids into host cells. Introducing recombinant plasmids into bacterial cells is called transformation. This is done by mixing the plasmids with the cells in a
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Molecular Genetics Techniques BIT 220 Chapter 20 What is Cloning? Recombinant DNA technologies Restriction Endonucleases Why use REs? Plasmids Cloning Vectors Plasmid Vector Other vectors Shuttle vectors Selection for E coli which contain plasmid Types of Genes No Lactose Present Lactose Present ...
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Summary Restriction sites and enzymes Cloning vectors (plasmids) Inserting foreign genes in plasmids Hosts cells for replicating plasmids (bacteria) Clone libraries, cDNA libraries Screening for recombinant plasmids Polymerase chain reaction ...
Date added: October 10, 2012 - Views: 39
Plasmids and other cloning vectors 7.1 DNA cloning with plasmid vectors Recombinant DNA technology depends on the ability to produce large numbers of identical DNA molecules (clones) Clones are typically generated by placing a DNA fragment of interest into a vector DNA molecule, ...
Date added: January 27, 2012 - Views: 11
Recombinant DNA Cont. Plasmids can carry up to 20,000 bp of foreign DNA. Besides bacterial plasmids, some other cloning vectors include viruses, bacteria artificial chromosomes (BACs), and yeast artificial chromosomes (YACs).
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Recombinant DNA cloning technology Chapter 7 DNA cloning To obtain large amounts of pure DNA Procedure Isolate DNA Use restriction enzymes to cut DNA Ligate fragments into a cloning vector Transform recombinant DNA into a host to replicate the DNA and pass copies into progeny.
Date added: October 8, 2011 - Views: 94
Plasmid Engineering p. 2-2 Transforming plasmids into bacteria p. 2-2 Safety Features Modern cloning plasmids have been ... MS Pゴシック Times Helvetica Symbol Blank Presentation Microsoft Word Document Slide 1 Introduction to Vectors Cloning Scheme Vector Types Plasmids ...
Date added: November 12, 2011 - Views: 93
Genomic libraries of higher organisms can be prepared by cloning M13 Vectors In the past, ... Contributions to plasmids Safety Features Modern cloning plasmids have been engineered so that they are incapable of transfer between bacterial cells Provide a level of biological containment.
Date added: May 20, 2013 - Views: 30
Plasmids as Cloning Vectors Plasmids are useful cloning vectors (Figure 10.36) because they are easy to isolate and purify and can multiply to high copy numbers in bacterial cells.
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Cloning Vectors. A. The DNA of interest is inserted into a cloning vector to transport it into the host cell. ... Engineered Ti plasmids lack the tumor-causing genes, but have the genes required to integrate the DNA of interest into the plant genome. (3) ...
Date added: May 26, 2012 - Views: 127
... .8 Figure 11.9 Figure 11.10 11.6 Plasmids as Cloning Vectors 11.6 Plasmids as Cloning Vectors Figure 11.11 11.6 Plasmids as Cloning Vectors Figure 11.12 11.7 Hosts for Cloning Vectors Figure 11.13 11.5 Gene Fusions and Reporter Genes 11.8 Shuttle Vectors and Expression Vectors ...
Date added: October 6, 2012 - Views: 55
Plasmid Cloning Vectors Plasmid cloning vectors have to be genetically engineered Naturally occurring plasmids often lacks important features such as Small size ... Plasmid Cloning Vectors Plasmids cloning vector A plasmid vector for cloning Slide 12 Slide 13 Slide 14 Continue!!
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... leaving sticky ends Artificial plasmids can be constructed by linking new DNA fragments to the sticky ends of plasmid Cloning Vectors Plasmids that can be modified to carry new genes Plasmids useful as cloning vectors must have a replicator (origin of replication) ...
Date added: March 3, 2012 - Views: 33
... is replicated and distributed along with plasmid Plasmid becomes a cloning vector delivers DNA into another cell Cloning Vectors Gene Libraries ... Engineering Discovery of Restriction Enzymes Specific Cuts Making Recombinant DNA Making Recombinant DNA Using Plasmids Cloning Vectors ...
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... DNA circles (only circles replicate in bacteria). Foreign DNA can be inserted into a plasmid and replicated. Plasmids for cloning carry drug resistance genes that are used for selection. ... Most cloning vectors use a system for detecting the presence of a recombinant insert, ...
Date added: May 8, 2012 - Views: 46
Recombinant DNA and Cloning Riyanda N G (10198) Vina E A (10221) Arini N (10268) Suluh N (10302) Cloning Definition The word "clone" has several different meanings in biology.
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Lecture # 04 Cloning Vectors Recombinant DNA Technology Common General Cloning Strategy Target DNA from donor organism extracted, cut with compatible restriction endonuclease and ligated into a cloning vector.
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Gene Cloning Part 2 Plasmid conformation Plasmid DNA may appear in one of five conformations, which (for a given size) run at different speeds in a gel during electrophoresis.
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Development of Plasmid Vectors Plasmids serve as important tools in genetics and biochemistry labs, where they are commonly used to multiply or express particular genes. ... In Molecular Cloning One of the major uses of plasmids is to make large amounts of proteins.
Date added: October 25, 2011 - Views: 46
... bacteria Recombinant plasmids Plasmids can be modified in biological labs Modified plasmid = Recombinant plasmid Plasmids can be used as cloning vectors to get the recombinant plasmid into E. coli Cloning vectors = way to get the gene of interest into the host Transformation Process in ...
Date added: April 1, 2013 - Views: 23
... so bacteria that aren’t transformed must be killed. Most cloning vectors have a multiple cloning site, a short region of DNA ... DNA circles (only circles replicate in bacteria). Foreign DNA can be inserted into a plasmid and replicated. Plasmids for cloning carry drug resistance ...
Date added: October 7, 2011 - Views: 240
... as transfer vehicles to insert DNA into cells Must be self-replicating and small enough to work with outside the cell Plasmids are common vectors ... What is Biotechnology Recombinant DNA How to make rDNA Slide 5 Slide 6 Restriction enzymes Restriction Enzymes Vectors Cloning ...
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Lambda vectors and their replication Sonita Gafary Biochem 72020 Lambda was first discovered at the Pasteur Institute by Andre Lwoff when he observed strains of E. Coli.
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H4 Eukaryotic vectors — Yeast episomal plasmids Vectors for the cloning and expression of genes in Saccharomyces cerevisiae. Based on 2 micron (2m) plasmid which is 6 kb in length. One origin Two genes ...
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... Bacteriophage λ Cosmid library Allows cloning of 45 kb DNA fragments BACs: ... the F plasmid and the lacZ region of pUC plasmids It’s a low copy number plasmid Carries 50-300kb ... cosmid vectors How is a Library Built: Restriction Enzyme Mechanisms: Preparation of DNAs to be ...
Date added: October 4, 2011 - Views: 149
Gene cloning Part 3 TYPES OF CLONING VECTORS BACTERIOPHAGE LAMBDA Phage lambda is a bacteriophage or phage, i.e. bacterial virus, that uses E. coli as host.
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... Cutting with EcoRI Cloning vectors can be: plasmids, bacteriophages, viruses, small artificial chromosomes Vectors have at least one unique cloning site: ...
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Composition Vectors Plasmids, or phage The cell E.coli, yeast Inserted sequence Vectors The substance that can serve as carriers to allow replication of recombinant DNAs. Plasmids Phage λ ... Three features of the plasmid cloning vectors: Multiple cloning site.
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Plasmids (Objectives of this Lecture) Understand the basic structures of plasmids Know the types of plasmids and their forms Undertand the methods used to transform E coli
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... vector Size of library depends on abundance of message Bacteriophage lambda insertion vectors or plasmids are used for cloning The choice depends upon: ... A plasmid vector for cloning Plasmid vectors Examples pBR322 One of the original plasmids used Two selectable markers (Amp and Tet ...
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Cloning vectors are plasmids that have been genetically engineered to reduce unneeded DNA and to introduce selectable markers such as antibiotic resistance genes (e.g., ampr) that are used to force cells to maintain the plasmid.
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However, R. plasmids can be useful vectors for genetic engineering. Plasmids are used to customize bacteria Page 234 1) Plasmid is isolated from bacterium 2) ... Cloning Genes in Recombinant Plasmids In order for recombinant DNA to be useful it must be made in vast amounts.
Date added: September 27, 2012 - Views: 10
Vectors for Gene Cloning Requirements of a vector to serve as a carrier molecule The choice of a vector depends on the design of the experimental ... vector Size of library depends on abundance of message Bacteriophage lambda insertion vectors or plasmids are often used for cloning cDNA ...
Date added: October 7, 2011 - Views: 102
Fact Sheet Describing Recombinant DNA and Elements Utilizing Recombinant DNA Such as Plasmids and Viral Vectors, and the Application of Recombinant DNA Techniques in ... Uses of recombinant DNA In vitro recombination Marker genes Transformation methods Cloning Vectors Plasmid vectors ...
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Cloning Vectors Cloning vectors are plasmids that can be modified to carry new genes Plasmids useful as cloning vectors must have: a replicator (origin of replication). a selectable marker (antibiotic resistance gene). a cloning site ...
Date added: August 23, 2011 - Views: 20
Chapter 13 An Introduction to Cloning and Recombinant DNA Clones Genetically identical organisms or molecules derived from a common ancestor Cloning Plants from Single Cells Fig. 13.1 Cloning Animals Animals were cloned more than 20 years ago Two techniques Embryo splitting Nuclear transfer ...
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Size of the insert DNA is limited in plasmid cloning vectors, and plasmids carrying more than 5–10 kb are often unstable. Fig. 8.5 Insertion of a piece of DNA into the plasmid cloning vector pUC19 Shuttle Vectors 1.
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Gene Cloning Techniques for gene ... complementary single-stranded stretches on other DNA molecules cut with the same restriction enzyme Recombinant DNA vectors Recombinant plasmids are produced by splicing restriction fragments from foreign DNA into plasmids.
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Learning Objectives Define Clone and DNA Cloning List the three steps of production of ... Lactose Metabolism plasmids and nucleic acid ... p. 375 Gene of interest DNA fragments with sticky ends Cell Restriction site Cut plasmid cloning vectors with a restriction enzyme to ...
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Cloning Vectors Enable DNA molecules to be replicated inside host (e.g., bacteria) cells. Features: Origin of replication (ORI) Cloning sites = Restriction enzyme recognition sites Selection = Identify bacteria that contain recombinant DNA (i.e., vector + insert) Types of cloning vectors ...
Date added: May 13, 2013 - Views: 16