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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: 37
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: 92
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 ...
Date added: October 16, 2011 - Views: 192
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 ...
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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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Promega Corporation Education Resources Unit 006 To determine whether your bacterial host cells have “taken up” your plasmid when you are cloning, most plasmids contain some sort of marker for antibiotic resistance.
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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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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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Cloning vectors “The introduction of a foreign DNA into a host cell in many cases requires the use of a vector. Vectors are DNA molecules used to transfer a gene into a host (microbial, plant, animal) cell, and to provide control elements for replication and expression.
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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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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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Molecular Cloning: Construction of a recombinant DNA Molecular Cloning: Cloning Vectors Plasmids Molecular Cloning: Cloning Vectors Plasmids Origin of replication, determines the number of copies per cell Marker genes: ampicillin and tetracycline resistance genes Unique restriction enzyme cut ...
Date added: May 6, 2013 - Views: 2
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: 100
3 Basic Principles of Recombinant DNA Technology. ... A. Bacterial Vectors. 1. Plasmids. 2. Bacteriophage. 3. Cosmids. B. Vectors for Other Organisms. 1. Yeast Artificial Chromosomes (YACs) 2. Bacterial Artificial Chromosomes (BACs) 3. Plant ... Cloning Vectors.
Date added: May 26, 2012 - Views: 141
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 ...
Date added: March 14, 2012 - Views: 69
... 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) ...
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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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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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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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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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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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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: 51
... Hamilton Smith studied how bacterium Haemophilus influenzae cut invading viral DNA Discovered that bacteria have an enzyme that chops up viral ... Engineering Discovery of Restriction Enzymes Specific Cuts Making Recombinant DNA Making Recombinant DNA Using Plasmids Cloning Vectors ...
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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: 20
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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Consequently, additional types of cloning vectors have been developed for cloning larger fragments of DNA ... Molecular Cloning I: Restriction Enzymes, PCR, Plasmids, and Gel Electrophoresis Author: Montarop Yamabhai Created Date: 7/18/2004 7:38:52 AM
Date added: October 7, 2011 - Views: 265
... cut directly across both strands leaving all dsDNA Cloning Vectors Plasmid Vectors Plasmid vector pBR322 B. Bacteriophage l Vectors C. Yeast Artificial Chromosomes as Vectors Yeast artificial chromosome ... Special Plasmids designed for cloning large DNA fragments (100-300 kbp) ...
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Plasmids useful as cloning vectors must have: An origin of replication. A selectable marker (antibiotic resistance gene, such as ampr and tetr). Multiple cloning site (MCS) (site where insertion of foreign DNA will not disrupt replication or inactivate essential markers).
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Chapter 4 Molecular Cloning Methods Introduction The significance of gene cloning To elucidate the structure and function of genes. i.e.: investigating hGH gene hGH gene: <10-6 of human genome Problem 1: need kilograms of human genome DNA for 1μg hGH gene Problem 2: how to ...
Date added: January 28, 2012 - Views: 161
Most cloning vectors have a multiple cloning site, a short region of DNA containing many restriction sites close ... SSR Example Cell-Based Molecular Cloning The original recombinant DNA technique: 1974 by Cohen ... The recombinant plasmids are transformed into E. coli using heat plus ...
Date added: October 7, 2011 - Views: 250
Genetic Engineering = inserting a foreign gene of interest ... 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 ...
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Biotechnology and Recombinant DNA Chapter 9 I. Learning Objectives Why genetic engineering? Cloning basics Beyond the basics Overview of DNA manipulation II.
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... .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 ...
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... 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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Cut with RE to confirm presence of foreign DNA. 10. Run on gel to identify recombinant plasmids. ... Presentation Restriction Enzymes PowerPoint Presentation PowerPoint Presentation Common Restriction Enzymes Cloning DNA in Plasmid Vectors PowerPoint Presentation Plasmid Used to Carry ...
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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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Types of cloning vectors 1. plasmids (about 20kb) 2. Bacteriophage (bacterial viruses), 30-50kb inserts 3. Cosmids (35-50kb insert) 4. BACs Use fertility F plasmid
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plasmids A plasmid is a circular, ... Luc The pGL2 Luciferase Reporter Vectors(a) ... CAT The pCAT®3-Basic Vector lacks eukaryotic promoter and enhancer sequences, allowing maximum flexibility in cloning putative regulatory sequences.
Date added: October 29, 2011 - Views: 172
Cloning Genes in Recombinant Plasmids In order for recombinant DNA to be useful it must be made in vast amounts. ... 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) ...
Date added: September 27, 2012 - Views: 13
... (YACs) Hybrid molecule containing components of yeast, protozoa and bacterial plasmids Yeast: ORI = ARS (autonomously replicating sequence ... Easy to be isolated from the host cell. A plasmid vector for cloning Plasmid vectors Examples pBR322 One of the original plasmids used ...
Date added: May 2, 2013 - Views: 29
Cloning and Sequencing Project overview Background Project will have you cloning the gene that codes for the enzyme glyceraldehyde-3-phosphate dehydrogenase (GAPDH) GAPDH is a housekeeping gene necessary for survival GAPDH is an enzyme that is crucial for glycolysis to occur Glycolysis GAPDH can ...
Date added: May 3, 2013 - Views: 30
Outline the procedures for producing plasmid and phage vectors. Isolation of vector and gene-source DNA - bacterial plasmids & foreign DNA with ... Explain how vectors are used in recombinant DNA technology. Cloning vector a DNA molecule that can carry foreign DNA into a cell and ...
Date added: May 2, 2013 - Views: 34
Title: Molecular Cloning I: Restriction Enzymes, PCR, Plasmids, and Gel Electrophoresis Author: Montarop Yamabhai Created Date: 7/18/2004 7:38:52 AM
Date added: September 27, 2011 - Views: 437
... Bacteriophage λ Cosmid library Allows cloning of 45 kb DNA ... the F plasmid and the lacZ region of pUC plasmids It’s a low copy number plasmid ... bacteriophage, phagemid, cosmid vectors cDNA Library: Stores a representation of the mRNAs expressed at a certain time or ...
Date added: March 2, 2014 - Views: 10
Cloning:Recombinant DNA Multistep Process . Produce fragments of DNA using enzymes that cut DNA at specific base sequences. . ... General PowerPoint Presentation Types of Vectors and DNA delivery systems Types of Vectors Plasmids Plasmid vectors need ...
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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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DNA Technology Chapter 13 Lab ... with the same restriction enzyme can bind together to form a new sequence of nucleotides Recognizes CTTAAG * * Cloning Vectors Cloning vector is a carrier that is ... isolate the DNA from human cells and plasmids from the bacteria Use restriction ...
Date added: September 13, 2013 - Views: 29