This document provides an overview of DNA cloning and recombinant DNA technology. It discusses how restriction enzymes are used to cut DNA into fragments that can be combined in vitro to form recombinant DNA molecules. Gene cloning involves inserting foreign DNA into a plasmid vector, which is then introduced into bacteria. The bacteria replicate the recombinant plasmid, producing multiple copies of the gene of interest. Libraries of cloned DNA fragments can be stored in bacteria or plasmids. Clones carrying a gene of interest can be identified by screening the library with a complementary DNA probe. Cloned genes can be expressed in bacterial or eukaryotic cells to produce the encoded protein.
1) Gene cloning involves inserting foreign DNA into plasmids which are then inserted into bacterial cells. The bacterial cells rapidly multiply, producing multiple copies of the gene of interest.
2) Restriction enzymes and DNA ligase are used to cut and join DNA fragments to create recombinant DNA. Probes are used to screen libraries and identify clones containing genes of interest.
3) Cloned genes can be expressed in bacterial or eukaryotic cells to produce large quantities of the gene's protein product. This allows researchers to study protein function.
The document discusses biotechnology principles and processes. It defines biotechnology as using living systems to develop useful products. The oldest forms of biotechnology included fermentation to produce wine and beer. Modern biotechnology relies on two key techniques: genetic engineering to alter DNA and introduce genes into host organisms, and sterile culture techniques to grow microbes and cells at large scales. Recombinant DNA techniques allow isolation of specific genes and insertion into vectors like plasmids for transfer into host cells. This allows cloning and multiplication of the target gene. Restriction enzymes, vectors, host cells, DNA ligase and polymerase enzymes are important tools that enable recombinant DNA technology.
Recombinant DNA technology involves combining DNA from two different organisms and inserting it into a host. This is done by using restriction enzymes to cut the DNA into fragments, which are then inserted into cloning vectors like plasmids, bacteriophages, or artificial chromosomes. The recombinant DNA is then inserted into a host organism using techniques like transformation or transfection. Gel electrophoresis can be used to analyze the results and identify successful recombinant clones. While cloning has potential medical applications, reproductive cloning of humans remains unsafe and controversial.
Biotechnology uses cells or their components to produce products. Recombinant DNA technology involves genetic engineering by inserting genes into cells to turn them into "factories" producing products. Key steps include using restriction enzymes to cut DNA for insertion into vectors like plasmids, transforming cells to take up the vector, and selecting clones containing the gene of interest, often using marker genes. Products can then be produced by growing the engineered cells in bacteria, yeast, or mammalian cells.
Recombinant DNA technology involves combining DNA from different sources and introducing it into a host cell. This allows for precise genetic analysis and practical applications. Key developments included elucidating DNA structure, cracking the genetic code, and describing transcription and translation. Gene cloning was developed in the 1970s, enabling previously impossible experiments. It involves isolating DNA, cutting it with restriction enzymes, ligating it into a vector, transforming host cells to amplify the recombinant DNA. The polymerase chain reaction (PCR) allows amplifying specific DNA regions without living cells by repeated heating and cooling in a test tube. It has revolutionized research fields like genetics and molecular biology.
Genetic modification through recombination breeding j.dJagdeep Singh
This document discusses genetic manipulation through recombinant breeding and various genetic engineering techniques. It defines genetic manipulation as the manipulation of genetic material to produce specific results in an organism. It then discusses recombinant DNA and various modern genetic modification techniques used, including Agrobacterium tumefaciens mediated transformation, biolistic methods, microinjection, electroporation, and lipofection. Examples of genetically engineered crops and their traits are provided. Both advantages and risks of genetic engineering are mentioned.
This presentation covers a general introduction to expression vector, its components, types, and its application. Then it covers some of the expression system with examples.
1) Gene cloning involves inserting foreign DNA into plasmids which are then inserted into bacterial cells. The bacterial cells rapidly multiply, producing multiple copies of the gene of interest.
2) Restriction enzymes and DNA ligase are used to cut and join DNA fragments to create recombinant DNA. Probes are used to screen libraries and identify clones containing genes of interest.
3) Cloned genes can be expressed in bacterial or eukaryotic cells to produce large quantities of the gene's protein product. This allows researchers to study protein function.
The document discusses biotechnology principles and processes. It defines biotechnology as using living systems to develop useful products. The oldest forms of biotechnology included fermentation to produce wine and beer. Modern biotechnology relies on two key techniques: genetic engineering to alter DNA and introduce genes into host organisms, and sterile culture techniques to grow microbes and cells at large scales. Recombinant DNA techniques allow isolation of specific genes and insertion into vectors like plasmids for transfer into host cells. This allows cloning and multiplication of the target gene. Restriction enzymes, vectors, host cells, DNA ligase and polymerase enzymes are important tools that enable recombinant DNA technology.
Recombinant DNA technology involves combining DNA from two different organisms and inserting it into a host. This is done by using restriction enzymes to cut the DNA into fragments, which are then inserted into cloning vectors like plasmids, bacteriophages, or artificial chromosomes. The recombinant DNA is then inserted into a host organism using techniques like transformation or transfection. Gel electrophoresis can be used to analyze the results and identify successful recombinant clones. While cloning has potential medical applications, reproductive cloning of humans remains unsafe and controversial.
Biotechnology uses cells or their components to produce products. Recombinant DNA technology involves genetic engineering by inserting genes into cells to turn them into "factories" producing products. Key steps include using restriction enzymes to cut DNA for insertion into vectors like plasmids, transforming cells to take up the vector, and selecting clones containing the gene of interest, often using marker genes. Products can then be produced by growing the engineered cells in bacteria, yeast, or mammalian cells.
Recombinant DNA technology involves combining DNA from different sources and introducing it into a host cell. This allows for precise genetic analysis and practical applications. Key developments included elucidating DNA structure, cracking the genetic code, and describing transcription and translation. Gene cloning was developed in the 1970s, enabling previously impossible experiments. It involves isolating DNA, cutting it with restriction enzymes, ligating it into a vector, transforming host cells to amplify the recombinant DNA. The polymerase chain reaction (PCR) allows amplifying specific DNA regions without living cells by repeated heating and cooling in a test tube. It has revolutionized research fields like genetics and molecular biology.
Genetic modification through recombination breeding j.dJagdeep Singh
This document discusses genetic manipulation through recombinant breeding and various genetic engineering techniques. It defines genetic manipulation as the manipulation of genetic material to produce specific results in an organism. It then discusses recombinant DNA and various modern genetic modification techniques used, including Agrobacterium tumefaciens mediated transformation, biolistic methods, microinjection, electroporation, and lipofection. Examples of genetically engineered crops and their traits are provided. Both advantages and risks of genetic engineering are mentioned.
This presentation covers a general introduction to expression vector, its components, types, and its application. Then it covers some of the expression system with examples.
Genetic transformation & success of DNA ligation Sabahat Ali
DNA is ligated through DNA Ligase, problems may occur during DNA ligation are
1) vector cyclization
2) vector-vector concatemers
3) target DNA-target DNA ligation
There are three main methods for isolating genes:
1. Using an automated gene machine to synthesize genes from predetermined nucleotide sequences.
2. Gene cloning, which involves inserting a DNA fragment into a vector that is then transferred into a host cell to produce multiple copies.
3. Polymerase chain reaction (PCR), which amplifies a specific DNA sequence using primers that flank the target sequence.
This document summarizes the structure and replication of genomes. It discusses the structure of prokaryotic and eukaryotic genomes, including bacterial chromosomes, plasmids, and eukaryotic nuclear and extranuclear DNA. It also describes DNA replication as semiconservative and requiring triphosphate deoxyribonucleotides. Key steps in DNA replication include initiation at the origin, replication by DNA polymerase in the 5' to 3' direction, and discontinuous lagging strand synthesis using Okazaki fragments.
This document summarizes DNA technology concepts from a biology textbook chapter. It discusses:
1) Advances in DNA sequencing techniques have allowed researchers to sequence the genomes of extinct species like Neanderthals and woolly mammoths. Next-generation sequencing is faster and less expensive than previous methods.
2) DNA cloning and sequencing are useful tools that have applications in fields like agriculture, criminal law, and medical research. DNA cloning involves inserting a DNA fragment into a plasmid vector, which is then replicated in bacteria to produce multiple copies of the fragment.
3) DNA sequencing determines the order of nucleotides in a gene or genome. The Sanger method uses DNA polymerase and dideoxynucleotides to generate
This document discusses two case studies involving genetically modified crops:
1) Drought tolerant transgenic plants developed using genes for abiotic stress tolerance through genetic engineering. Genes used include structural genes for late embryogenesis abundant (LEA) proteins and regulatory genes for key enzymes. This allows improved water stress management compared to conventional breeding.
2) Genetically engineered potatoes (Innate) modified using RNA interference to suppress expression of certain genes to reduce browning and increase resistance to bruising and soft rot. This helps improve potato quality and shelf life.
This document provides instructions for preparing competent bacterial cells for transformation. Key steps include growing cells to mid-log phase, treating with calcium chloride to make membranes permeable, adding exogenous DNA, and applying a heat shock to induce DNA uptake. The resulting transformed competent cells can be used to take up exogenous DNA from their surroundings through their membranes. Proper preparation is important for achieving high transformation efficiency.
Cloning involves making genetically identical copies of DNA, cells, or organisms through asexual reproduction. There are two main methods for cloning DNA - recombinant DNA technology and polymerase chain reaction. Recombinant DNA technology involves isolating a gene, inserting it into a plasmid, and transforming the plasmid into a host cell to mass produce the gene. Polymerase chain reaction allows scientists to make copies of available DNA. Today, transgenic bacteria, plants, and animals produce useful biotechnology products through genetic engineering.
Recombinant DNA technology involves isolating a gene of interest, inserting it into a vector, transferring the recombinant DNA into a host cell, and identifying cells that contain the recombinant DNA. Key steps include isolating the target DNA through restriction enzyme digestion or PCR, selecting a vector like a plasmid or phage, ligating the DNA insert into the vector, transforming host cells, and using selection methods like antibiotic resistance to identify recombinant cells. This allows large scale production and study of the target gene.
This document discusses cell transformation, which is a change to a cell's DNA. It describes three methods of transforming cells:
1) Transforming bacteria cells using plasmids containing foreign DNA that can be inserted into the plasmid. The recombinant plasmids can then deliver the DNA into the bacterial cell.
2) Transforming plant cells either by using the bacterium Agrobacterium tumefaciens to transfer recombinant plasmids, or by using a gene gun to inject DNA-coated particles into plant cells.
3) Transforming animal cells by injecting foreign DNA directly into egg nuclei, or using gene replacement to "knock out" an existing gene.
Recombinant DNA (rDNA) refers to DNA created outside living cells by joining DNA from multiple sources. Common techniques for creating rDNA include restriction enzymes to cut DNA strands, ligation to join strands, and transformation or transfection to introduce rDNA into host cells. Vectors like plasmids, viruses, and artificial chromosomes are often used to replicate and express rDNA in host cells. rDNA techniques have applications in gene cloning, DNA sequencing, genetic engineering of plants and animals, and gene therapy to treat diseases.
Vectors are used to carry DNA fragments into host cells for replication. Plasmids are commonly used cloning vectors that are extrachromosomal and autonomously replicating in bacteria. Properties of good vectors include an origin of replication, antibiotic resistance marker, and unique restriction sites. Different vector types include plasmids, bacteriophages, cosmids, BACs, YACs, and mini chromosomes, each with advantages for cloning varying size DNA fragments. Recombinant DNA technology uses restriction enzymes to cut DNA, ligase to join fragments, and vectors to replicate, express, and select for cloned genes.
A genetically engineered DNA molecule from bacteria , phage or yeast to carry foreign DNA for the purpose of cloning and expression of the inserted DNA of interest in RDT
Recombinant DNA technology involves transferring genes between organisms using artificial means. It works by combining DNA from different sources into a single molecule. The process involves generating DNA fragments, inserting the fragments into vectors, introducing the vectors into host cells, and selecting clones containing the recombinant DNA. Common tools used include restriction enzymes to cut DNA, vectors like plasmids to carry DNA, bacterial hosts like E. coli, and techniques like transformation and selection to introduce and identify recombinant DNA. Applications include analyzing gene structure, producing pharmaceuticals, genetically modified organisms, and gene therapy.
This document discusses several applications of biotechnology including gene cloning, DNA fingerprinting, and genetically modified foods. It provides information on how restriction enzymes can cut DNA at specific sequences to create fragments for gene cloning. DNA fragments can be joined together to form recombinant DNA which is often inserted into plasmids replicated in bacteria. This allows production of human insulin by bacteria. DNA fingerprinting analyzes variable regions of DNA to produce unique patterns that can be used for identification. While genetically modified crops are widely grown, scientists are still examining their potential environmental impacts.
Recombinant DNA technology involves isolating DNA from different species, cutting it with restriction enzymes, and splicing the pieces together to form new recombinant molecules. These molecules are then inserted into host cells like bacteria or yeast where they can be replicated in large quantities. Key aspects of the process include using restriction enzymes to cut DNA at specific recognition sequences, producing DNA fragments with cohesive or blunt ends, and inserting the fragments into plasmids - small extrachromosomal DNA molecules found in bacteria. Plasmids are often used as vectors to carry foreign DNA, and they allow selection of cells containing the recombinant DNA through the use of antibiotic resistance genes on the plasmid.
The document discusses biotechnology and recombinant DNA technology. It defines biotechnology as using organisms or enzymes from organisms to produce useful products. Recombinant DNA technology involves isolating DNA, fragmenting it with enzymes, inserting fragments into vectors, transforming host cells, and culturing the cells to multiply the DNA. The basic steps are isolating a gene, inserting it into a vector, introducing the vector into a host cell, and using the host to generate multiple copies of the gene.
Recombinant DNA technology allows DNA from different species to be isolated, cut with restriction enzymes, and spliced together to form new recombinant molecules. This involves extracting DNA, cutting it with restriction enzymes to form manageable fragments, inserting fragments into vectors like plasmids, introducing the recombinant vectors into host cells, and amplifying the DNA. Vectors often contain antibiotic resistance genes to select for host cells containing the recombinant DNA. This process allows scientists to isolate and multiply specific genes for study and modification.
Gene cloning involves inserting foreign DNA into bacterial plasmids. Restriction enzymes cut DNA at specific sites, creating sticky or blunt ends. Plasmids can replicate independently and accept foreign DNA fragments. The recombinant plasmid is inserted into bacteria via transformation or electroporation. Transformed bacteria are selected using antibiotics or blue-white screening to identify those containing the recombinant plasmid, allowing mass production of the cloned gene.
bacteriophages require bacterial host to complete its life-cycle, wherein site-specific genetic recombination occurs. furthermore, homologous recombination also occur in phages in case of multiple infection of the host cell.
Gene cloning involves inserting DNA fragments from one source into plasmids or bacteria. This allows for the production of multiple copies of the gene. Restriction enzymes and DNA ligase are used to cut and paste DNA fragments into plasmids. The plasmids are then inserted into bacteria which replicate, producing many copies of the gene. Libraries of cloned DNA fragments can be stored in bacteria or plasmids. The library can be screened to identify clones containing genes of interest using probes complementary to the target gene. Expressed genes can be studied by producing their protein products in bacterial or eukaryotic cells.
The document outlines the major discoveries in DNA research from 1910 to 1953. It begins with Thomas Morgan's confirmation in 1910 that chromosomes are the molecules of inheritance. Important early experiments were conducted by Griffith in 1928 and Avery in 1944 suggesting DNA was the molecule of inheritance. In 1950, Chargaff determined purine and pyrimidine amounts were equal in DNA strands. Franklin photographed DNA in 1952 providing evidence it was a double strand. Watson and Crick used this photo and Chargaff's rules to develop their 1953 double helix model of DNA's structure.
Genetic transformation & success of DNA ligation Sabahat Ali
DNA is ligated through DNA Ligase, problems may occur during DNA ligation are
1) vector cyclization
2) vector-vector concatemers
3) target DNA-target DNA ligation
There are three main methods for isolating genes:
1. Using an automated gene machine to synthesize genes from predetermined nucleotide sequences.
2. Gene cloning, which involves inserting a DNA fragment into a vector that is then transferred into a host cell to produce multiple copies.
3. Polymerase chain reaction (PCR), which amplifies a specific DNA sequence using primers that flank the target sequence.
This document summarizes the structure and replication of genomes. It discusses the structure of prokaryotic and eukaryotic genomes, including bacterial chromosomes, plasmids, and eukaryotic nuclear and extranuclear DNA. It also describes DNA replication as semiconservative and requiring triphosphate deoxyribonucleotides. Key steps in DNA replication include initiation at the origin, replication by DNA polymerase in the 5' to 3' direction, and discontinuous lagging strand synthesis using Okazaki fragments.
This document summarizes DNA technology concepts from a biology textbook chapter. It discusses:
1) Advances in DNA sequencing techniques have allowed researchers to sequence the genomes of extinct species like Neanderthals and woolly mammoths. Next-generation sequencing is faster and less expensive than previous methods.
2) DNA cloning and sequencing are useful tools that have applications in fields like agriculture, criminal law, and medical research. DNA cloning involves inserting a DNA fragment into a plasmid vector, which is then replicated in bacteria to produce multiple copies of the fragment.
3) DNA sequencing determines the order of nucleotides in a gene or genome. The Sanger method uses DNA polymerase and dideoxynucleotides to generate
This document discusses two case studies involving genetically modified crops:
1) Drought tolerant transgenic plants developed using genes for abiotic stress tolerance through genetic engineering. Genes used include structural genes for late embryogenesis abundant (LEA) proteins and regulatory genes for key enzymes. This allows improved water stress management compared to conventional breeding.
2) Genetically engineered potatoes (Innate) modified using RNA interference to suppress expression of certain genes to reduce browning and increase resistance to bruising and soft rot. This helps improve potato quality and shelf life.
This document provides instructions for preparing competent bacterial cells for transformation. Key steps include growing cells to mid-log phase, treating with calcium chloride to make membranes permeable, adding exogenous DNA, and applying a heat shock to induce DNA uptake. The resulting transformed competent cells can be used to take up exogenous DNA from their surroundings through their membranes. Proper preparation is important for achieving high transformation efficiency.
Cloning involves making genetically identical copies of DNA, cells, or organisms through asexual reproduction. There are two main methods for cloning DNA - recombinant DNA technology and polymerase chain reaction. Recombinant DNA technology involves isolating a gene, inserting it into a plasmid, and transforming the plasmid into a host cell to mass produce the gene. Polymerase chain reaction allows scientists to make copies of available DNA. Today, transgenic bacteria, plants, and animals produce useful biotechnology products through genetic engineering.
Recombinant DNA technology involves isolating a gene of interest, inserting it into a vector, transferring the recombinant DNA into a host cell, and identifying cells that contain the recombinant DNA. Key steps include isolating the target DNA through restriction enzyme digestion or PCR, selecting a vector like a plasmid or phage, ligating the DNA insert into the vector, transforming host cells, and using selection methods like antibiotic resistance to identify recombinant cells. This allows large scale production and study of the target gene.
This document discusses cell transformation, which is a change to a cell's DNA. It describes three methods of transforming cells:
1) Transforming bacteria cells using plasmids containing foreign DNA that can be inserted into the plasmid. The recombinant plasmids can then deliver the DNA into the bacterial cell.
2) Transforming plant cells either by using the bacterium Agrobacterium tumefaciens to transfer recombinant plasmids, or by using a gene gun to inject DNA-coated particles into plant cells.
3) Transforming animal cells by injecting foreign DNA directly into egg nuclei, or using gene replacement to "knock out" an existing gene.
Recombinant DNA (rDNA) refers to DNA created outside living cells by joining DNA from multiple sources. Common techniques for creating rDNA include restriction enzymes to cut DNA strands, ligation to join strands, and transformation or transfection to introduce rDNA into host cells. Vectors like plasmids, viruses, and artificial chromosomes are often used to replicate and express rDNA in host cells. rDNA techniques have applications in gene cloning, DNA sequencing, genetic engineering of plants and animals, and gene therapy to treat diseases.
Vectors are used to carry DNA fragments into host cells for replication. Plasmids are commonly used cloning vectors that are extrachromosomal and autonomously replicating in bacteria. Properties of good vectors include an origin of replication, antibiotic resistance marker, and unique restriction sites. Different vector types include plasmids, bacteriophages, cosmids, BACs, YACs, and mini chromosomes, each with advantages for cloning varying size DNA fragments. Recombinant DNA technology uses restriction enzymes to cut DNA, ligase to join fragments, and vectors to replicate, express, and select for cloned genes.
A genetically engineered DNA molecule from bacteria , phage or yeast to carry foreign DNA for the purpose of cloning and expression of the inserted DNA of interest in RDT
Recombinant DNA technology involves transferring genes between organisms using artificial means. It works by combining DNA from different sources into a single molecule. The process involves generating DNA fragments, inserting the fragments into vectors, introducing the vectors into host cells, and selecting clones containing the recombinant DNA. Common tools used include restriction enzymes to cut DNA, vectors like plasmids to carry DNA, bacterial hosts like E. coli, and techniques like transformation and selection to introduce and identify recombinant DNA. Applications include analyzing gene structure, producing pharmaceuticals, genetically modified organisms, and gene therapy.
This document discusses several applications of biotechnology including gene cloning, DNA fingerprinting, and genetically modified foods. It provides information on how restriction enzymes can cut DNA at specific sequences to create fragments for gene cloning. DNA fragments can be joined together to form recombinant DNA which is often inserted into plasmids replicated in bacteria. This allows production of human insulin by bacteria. DNA fingerprinting analyzes variable regions of DNA to produce unique patterns that can be used for identification. While genetically modified crops are widely grown, scientists are still examining their potential environmental impacts.
Recombinant DNA technology involves isolating DNA from different species, cutting it with restriction enzymes, and splicing the pieces together to form new recombinant molecules. These molecules are then inserted into host cells like bacteria or yeast where they can be replicated in large quantities. Key aspects of the process include using restriction enzymes to cut DNA at specific recognition sequences, producing DNA fragments with cohesive or blunt ends, and inserting the fragments into plasmids - small extrachromosomal DNA molecules found in bacteria. Plasmids are often used as vectors to carry foreign DNA, and they allow selection of cells containing the recombinant DNA through the use of antibiotic resistance genes on the plasmid.
The document discusses biotechnology and recombinant DNA technology. It defines biotechnology as using organisms or enzymes from organisms to produce useful products. Recombinant DNA technology involves isolating DNA, fragmenting it with enzymes, inserting fragments into vectors, transforming host cells, and culturing the cells to multiply the DNA. The basic steps are isolating a gene, inserting it into a vector, introducing the vector into a host cell, and using the host to generate multiple copies of the gene.
Recombinant DNA technology allows DNA from different species to be isolated, cut with restriction enzymes, and spliced together to form new recombinant molecules. This involves extracting DNA, cutting it with restriction enzymes to form manageable fragments, inserting fragments into vectors like plasmids, introducing the recombinant vectors into host cells, and amplifying the DNA. Vectors often contain antibiotic resistance genes to select for host cells containing the recombinant DNA. This process allows scientists to isolate and multiply specific genes for study and modification.
Gene cloning involves inserting foreign DNA into bacterial plasmids. Restriction enzymes cut DNA at specific sites, creating sticky or blunt ends. Plasmids can replicate independently and accept foreign DNA fragments. The recombinant plasmid is inserted into bacteria via transformation or electroporation. Transformed bacteria are selected using antibiotics or blue-white screening to identify those containing the recombinant plasmid, allowing mass production of the cloned gene.
bacteriophages require bacterial host to complete its life-cycle, wherein site-specific genetic recombination occurs. furthermore, homologous recombination also occur in phages in case of multiple infection of the host cell.
Gene cloning involves inserting DNA fragments from one source into plasmids or bacteria. This allows for the production of multiple copies of the gene. Restriction enzymes and DNA ligase are used to cut and paste DNA fragments into plasmids. The plasmids are then inserted into bacteria which replicate, producing many copies of the gene. Libraries of cloned DNA fragments can be stored in bacteria or plasmids. The library can be screened to identify clones containing genes of interest using probes complementary to the target gene. Expressed genes can be studied by producing their protein products in bacterial or eukaryotic cells.
The document outlines the major discoveries in DNA research from 1910 to 1953. It begins with Thomas Morgan's confirmation in 1910 that chromosomes are the molecules of inheritance. Important early experiments were conducted by Griffith in 1928 and Avery in 1944 suggesting DNA was the molecule of inheritance. In 1950, Chargaff determined purine and pyrimidine amounts were equal in DNA strands. Franklin photographed DNA in 1952 providing evidence it was a double strand. Watson and Crick used this photo and Chargaff's rules to develop their 1953 double helix model of DNA's structure.
When DNA adopts an unusual Z-shaped structure, it can lead to genetic instability and breaks in the DNA that have been associated with cancers like leukemia. A study showed that the unusual Z-DNA shape can directly cause breaks in mammalian cell DNA. Regions of DNA prone to forming Z-DNA are also often genetic "hot spots" with high rates of rearrangement associated with cancer. The odd Z-DNA shape may cause the DNA molecule to completely break apart.
The document provides an overview of environmental policy, law, and planning. It discusses major environmental laws in the US like the Clean Air Act, Clean Water Act, and Endangered Species Act. It also covers how environmental policies are made through statutes, case law, and administrative rules. Regulatory agencies implement policies at the federal, state, and local levels. Public participation, lobbying, and international treaties also influence environmental governance.
The document discusses the Polymerase Chain Reaction (PCR) technique. It begins by explaining that PCR amplifies DNA fragments rapidly outside of a cell. It then notes that PCR was invented in 1984 as a way to make numerous copies of DNA fragments in the laboratory. The document proceeds to describe the components needed for PCR, including DNA, primers, nucleotides, DNA polymerase, and thermal cyclers. It explains the three step PCR process of denaturation, annealing of primers, and extension of DNA. Finally, it outlines several applications and variants of the PCR technique.
Concept: reannealing nucleic acids to identify sequence of interest.
Separates DNA/RNA in an agarose gel, then detects specific bands using probe and hybridization.
Hybridization takes advantage of the ability of a single stranded DNA or RNA molecule to find its complement, even in the presence of large amounts of unrelated DNA.
Allows detection of specific bands (DNA fragments or RNA molecules) that have complementary sequence to the probe.
Size bands and quantify abundance of molecule.
Polymerase chain reaction (PCR) is a technique used to amplify DNA sequences. It requires a DNA template, primers, dNTPs, buffer/magnesium, and DNA polymerase. Each PCR cycle involves denaturation of the DNA, annealing of primers, and extension of the DNA strands. Multiple cycles are done to exponentially amplify the target sequence. PCR is widely used in research, forensics, medicine, and many other fields.
The document compares the relative sizes of subcellular structures using SI units. It states that plant cells are 100μm, animal cells are 10μm, bacteria are 1μm, and viruses are between 50-100nm. It also discusses calculating linear magnification by taking the ratio of image size to real size. Finally, it states that understanding individual parts of multicellular organisms does not allow understanding of emergent properties that arise from their interaction.
The document summarizes the Southern blot technique. It involves digesting DNA with restriction enzymes, separating fragments by size via gel electrophoresis, transferring fragments to a membrane, and detecting a specific DNA sequence using a radiolabeled probe. The Southern blot allows determining the size of a restriction fragment, measuring amounts between samples, and locating a specific sequence within a complex DNA mixture. It has applications in gene discovery, mapping, identification of transgenic genes, and DNA fingerprinting.
The document describes regulation of gene expression in prokaryotic and eukaryotic cells. In prokaryotes, gene expression is regulated through operons, clusters of genes that are coordinately controlled. The lac and trp operons in E. coli are discussed as examples, with the lac operon being inducible and the trp operon being repressible. In eukaryotes, gene expression can be regulated at many stages including chromatin modifications, transcription, RNA processing, translation and protein modification. This allows for cell specialization and differential gene expression with the same genome.
The document discusses the history and development of the polymerase chain reaction (PCR) technique. It describes how Kary Mullis invented PCR in 1985 and was awarded the Nobel Prize for it. It then explains the basic steps of PCR including denaturation, annealing of primers, and extension. Finally, it discusses several variations and applications of PCR including real-time PCR, asymmetric PCR, and comparisons to cloning techniques.
Gene cloning allows scientists to produce multiple copies of a gene or DNA segment. The process involves using restriction enzymes to cut DNA into fragments, which are then combined with a bacterial plasmid. This recombinant DNA is inserted into bacterial cells via transformation. The bacteria are then cultured to generate clones containing many copies of the gene of interest. DNA cloning has many applications, including producing proteins for research and modifying organisms.
1) Gene cloning involves inserting foreign DNA into a plasmid, which is then inserted into bacterial cells. The bacterial cells rapidly replicate the recombinant plasmid, producing multiple copies of the gene of interest.
2) Restriction enzymes are used to cut DNA at specific sites, producing sticky ends that allow insertion of the foreign DNA fragment into the plasmid. DNA ligase seals the recombinant DNA molecule.
3) The recombinant plasmids are inserted into bacteria, which are then cultured to produce clones containing many copies of the cloned gene, allowing mass production of that gene or its protein product.
1) The document describes techniques for cloning DNA fragments, including using restriction enzymes to cut DNA into fragments and recombining fragments into plasmids, which are then inserted into bacterial cells.
2) The cloned DNA fragments can be used to create libraries that allow screening for genes of interest. Once identified, the gene can be isolated and studied.
3) DNA technology allows researchers to analyze genes through techniques like PCR, gel electrophoresis, and Southern blotting to compare sequences, locate expression, and determine function.
The document discusses DNA technology and gene cloning. It describes how the human genome was sequenced by 2003 through advances in DNA technology including the invention of recombinant DNA methods. Gene cloning allows scientists to prepare multiple identical copies of a specific gene or DNA segment. This involves inserting the gene into a bacterial plasmid, which is then put into bacterial cells to produce clones containing the gene of interest. Restriction enzymes and DNA ligase are used to create recombinant DNA by cutting DNA into fragments that can be recombined. The polymerase chain reaction can amplify a specific DNA sequence in vitro. Gel electrophoresis and Southern blotting allow researchers to analyze and compare DNA restriction fragments.
1) Gene cloning involves inserting foreign DNA into a plasmid vector, transforming bacteria with the recombinant plasmid, and allowing the bacteria to replicate, producing multiple copies of the gene of interest.
2) Restriction enzymes are used to cut the DNA at specific sites, producing sticky ends that allow insertion of the gene into the plasmid.
3) Transformed bacteria containing the recombinant plasmid are selected by growing on antibiotic-containing media, with the antibiotic resistance gene on the plasmid allowing only those bacteria to survive.
This document provides an overview of DNA cloning including:
1. The basic steps in DNA cloning including isolation of vector and gene source DNA, insertion into the vector, and introduction into cells.
2. Uses of polymerase chain reaction and restriction enzymes in cloning.
3. Applications of cloning such as recombinant protein production, genetically modified organisms, DNA fingerprinting, and gene therapy.
Genetic engineering involves directly manipulating genes, often by adding a gene from another species to an organism's genome. This is done through recombinant DNA (rDNA) technology, which combines DNA sequences artificially. A key part of the process is using restriction enzymes to cut DNA at specific sites, then inserting the cut DNA fragment into a vector like a plasmid for replication in a host cell. The engineered DNA is then introduced into host cells, and cells containing the new DNA are identified and isolated through markers on the vector.
This document discusses various methods and tools for gene cloning, including:
- Plasmids as cloning vectors, with examples like pUC19, which contains features like antibiotic resistance markers and multiple cloning sites.
- Host cells used for cloning vectors, including E. coli, yeast, and other prokaryotic and eukaryotic cells.
- Shuttle vectors that can replicate in multiple host organisms, and expression vectors that control gene expression.
- Bacteriophage lambda and modified lambda phages like Charon that can be used as cloning vectors for larger DNA fragments.
- Additional tools for cloning genes into vectors, expressing genes, and transferring vectors between prokaryotic and eukaryotic hosts
The procedure involves placing a foreign gene into bacterial cells using restriction enzymes and vectors. The modified bacteria are then grown to produce clones. Key aspects of gene cloning include restriction endonucleases to cut DNA, vectors to allow DNA replication, and probes to identify specific clones. DNA ligase is used to join cut DNA fragments.
Biotechnology refers to the use of living organisms or their components to develop products and processes. It has applications in fields like agriculture, medicine, and industry. Modern biotechnology techniques include genetic engineering and aseptic techniques. Genetic engineering involves altering genetic material through techniques like recombinant DNA, gene transfer into host organisms, and gene cloning. It allows scientists to modify organisms for useful purposes. Restriction enzymes, vectors, DNA polymerase and ligase are important tools used in genetic engineering and recombinant DNA technology.
This document provides an overview of DNA cloning. It begins by defining cloning as making identical copies of DNA, genes, or cells. The basic steps of DNA cloning are described, including using a source DNA, vector, restriction enzymes to cut DNA, ligation to join DNA fragments, transformation of host bacteria, and selection of recombinant clones. Common vectors like plasmids are discussed along with selection techniques like blue-white screening. The document emphasizes that the goal is to generate multiple copies of the cloned insert DNA. Examples are given of important medical and agricultural applications of cloning genes.
1. The document discusses genetic engineering and biotechnology, including techniques like recombinant DNA, gene cloning, PCR, and cloning organisms.
2. Key terms defined include genetic engineering, biotechnology, recombinant DNA, gene cloning, restriction enzymes, and plasmids.
3. Recombinant DNA is made by cutting DNA with restriction enzymes and ligating pieces into plasmids, which are then inserted into bacteria.
This document provides an overview of DNA cloning. It discusses taking a gene of interest from a source DNA, inserting it into a vector such as a plasmid, and using this recombinant DNA to transform bacteria. This allows the gene of interest to be replicated in large quantities. Key steps include using restriction enzymes to cut the DNA pieces for ligation, transforming bacteria with the recombinant plasmid, and selecting for bacteria containing the cloned gene insert. The goal of cloning is to generate multiple copies of a gene for study and protein production.
This document discusses recombinant DNA technology and its applications. It begins with an introduction to recombinant DNA technology and its history. It then describes the tools and enzymes used, including restriction enzymes, DNA ligase, reverse transcriptase, and DNA polymerase. Various vectors like bacterial plasmids and bacteriophages are also discussed. The document outlines several applications such as monoclonal antibody production, disease diagnosis, DNA fingerprinting, environmental uses, gene therapy, and xenotransplantation. In summary, the document provides an overview of the key concepts, techniques, and uses of recombinant DNA technology.
CONFERENCE 5-Techniques in Genetic Engineering.pptDicksonDaniel7
This document describes genetic engineering techniques such as selective breeding, recombinant DNA, PCR, and transgenic organisms. It explains how recombinant DNA technology uses restriction enzymes and DNA ligases to insert DNA fragments into cloning vectors, which are then inserted into host bacteria for replication. This allows mass production of useful proteins like insulin. The document also discusses how Agrobacterium tumefaciens and its Ti plasmid are widely used vectors to introduce foreign DNA into plant cells and genomes. In summary, the document outlines genetic engineering methods and how they have been applied to biotechnology and agriculture.
Recombinant DNA technology allows DNA from different sources to be combined through molecular biology techniques. DNA segments are recombined outside of living cells and can then enter a host cell and replicate. This technology was developed in 1973 and allows genes to be transferred between organisms. Recombinant DNA is made through various methods like transformation, phage introduction, or non-bacterial transformation which insert DNA into vectors that are then taken up by host cells. This technology is used to produce human proteins like insulin in bacteria for medical purposes. Safety issues involve ensuring recombinant DNA does not escape the laboratory.
Genetic engineering involves transferring genes between organisms using recombinant DNA techniques. This allows genes to be isolated, cloned, and moved within and between different species. Cloning a gene involves using restriction enzymes to cut DNA at specific sequences, and DNA ligase to join DNA fragments together. Cloned genes have many research uses such as determining gene sequences, altering phenotypes, and obtaining protein products of genes.
Genetic engineering involves transferring genes between organisms using recombinant DNA techniques. This allows genes to be isolated, cloned, and moved within and between different species. Cloning a gene involves using restriction enzymes to cut DNA at specific sequences, and DNA ligase to join DNA fragments together. Cloned genes have many research uses such as determining gene sequences, altering phenotypes, and obtaining protein products of genes.
The document provides an overview of DNA technology and biotechnology. It discusses how DNA cloning allows scientists to make multiple copies of genes and study their structure, expression, and function. Key techniques described include recombinant DNA, restriction enzymes, gel electrophoresis, and DNA sequencing. Applications mentioned include genetic engineering of plants, microorganisms, and animals for research, agriculture, medicine, forensics, and environmental cleanup.
Recombinant DNA technology involves isolating DNA from different species, cutting it with restriction enzymes, and splicing the pieces together to form recombinant molecules. These molecules are multiplied in bacteria or yeast cells. Key steps include extracting DNA, cutting it with restriction enzymes, inserting a gene of interest into a plasmid, transforming bacteria with the plasmid, and using antibiotics to select bacteria containing the recombinant DNA. This allows mass production of human genes for applications like gene therapy and production of therapeutic proteins.
1. The document discusses various topics related to biotechnology and genetic engineering including organic compounds, bioprocessing techniques, tools and applications of genetic engineering.
2. Specific topics covered include microbial fermentation, genetic engineering of plants, animals and microbes, biomanufacturing, biosensors, bioremediation, tissue engineering and gene therapy.
3. The document provides information on biotechnology tools like enzymes, plasmids, vaccines and antibodies and their uses in areas such as biomanufacturing, diagnostics, environmental monitoring, pollution control, and disease prevention.
This document discusses food biotechnology and its past, present, and future applications. It defines food biotechnology as using modern genetics tools to enhance beneficial plant, animal, and microorganism traits for food production. Examples of benefits include reducing pesticide use, increasing crop yields, improving nutrition, and developing hardier plants. The document also examines consumer and regulatory perspectives, finding that most support food biotechnology and current FDA labeling policies. It predicts future applications could lower natural toxins and allergens while extending freshness and farming efficiency.
This document provides an introduction to biotechnology, describing it as using scientific processes to develop new organisms or products from organisms to meet human needs like food, clothing, shelter, health and safety. It discusses areas of biotechnology like agriculture, medicine, environment, and food/beverage processing. Key applications mentioned include genetically modified crops, animal biotechnology, gene therapy, biopharmaceuticals, and synthetic biology.
This document discusses plant biotechnology techniques used to genetically modify organisms. It defines biotechnology as applying technology to modify biological organisms by adding genes from other species. The key techniques discussed are identifying genes from other organisms that control desired traits and introducing those genes into plants through transformation. This allows developing crops with improved traits like herbicide or insect resistance, drought tolerance, or increased nutritional content. The document outlines the process of gene cloning, creation of transformation cassettes containing the gene of interest and selectable marker, and delivery into plants via Agrobacterium or gene gun. Extensive testing of transgenic plants in the lab and field is needed before commercial release to ensure safety and trait expression.
The document provides an overview of the pharmaceutical and biotechnology industries. It discusses how pharmaceutical companies produce drugs and other products, and how biotechnology companies use genetic research to develop products. It outlines trends in industry consolidation and partnerships between large pharmaceutical companies and smaller biotech firms. It also describes various career opportunities and paths within these industries.
Biotechnology is the use of living organisms or substances from living organisms to develop products or modify organisms. It involves techniques like genetic engineering and recombinant DNA technology. Some key areas of biotechnology include medicine, agriculture, food production, and industry. The field is multidisciplinary, drawing from areas like life sciences, physical sciences, engineering, and mathematics. Biotechnology has contributed to advances in areas like disease treatment, agriculture, and environmental management. It continues to be an area of growing career opportunities.
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আমাদের সবার জন্য খুব খুব গুরুত্বপূর্ণ একটি বই ..বিসিএস, ব্যাংক, ইউনিভার্সিটি ভর্তি ও যে কোন প্রতিযোগিতা মূলক পরীক্ষার জন্য এর খুব ইম্পরট্যান্ট একটি বিষয় ...তাছাড়া বাংলাদেশের সাম্প্রতিক যে কোন ডাটা বা তথ্য এই বইতে পাবেন ...
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Physiology and chemistry of skin and pigmentation, hairs, scalp, lips and nail, Cleansing cream, Lotions, Face powders, Face packs, Lipsticks, Bath products, soaps and baby product,
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8. 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
DNA of
chromosome
(“foreign” DNA)
Recombinant
bacterium
Host cell grown in culture to
form a clone of cells containing
the “cloned” gene of interest
Gene of
interest
Protein expressed from
gene of interest
Protein harvestedCopies of gene
Basic research
and various
applications
Basic
research
on protein
Basic
research
on gene
Gene for pest
resistance inserted
into plants
Gene used to alter
bacteria for cleaning
up toxic waste
Protein dissolves
blood clots in heart
attack therapy
Human growth
hormone treats
stunted growth
9. Figure 20.2a
Bacterium
Bacterial
chromosome
Plasmid
2
1 Gene inserted into
plasmid
Cell containing
gene of interest
Recombinant
DNA (plasmid)
Gene of
interest
Plasmid put into
bacterial cell
DNA of
chromosome
(“foreign” DNA)
Recombinant
bacterium
10. Figure 20.2b
Host cell grown in
culture to form a clone
of cells containing the
“cloned” gene of interest
Gene of
interest
Protein expressed from
gene of interest
Protein harvestedCopies of gene
Basic research
and various
applications
3
4
Basic
research
on protein
Basic
research
on gene
Gene for pest
resistance inserted
into plants
Gene used to alter
bacteria for cleaning
up toxic waste
Protein dissolves
blood clots in heart
attack therapy
Human growth
hormone treats
stunted growth
14. Figure 20.3-1
Restriction enzyme
cuts sugar-phosphate
backbones.
Restriction site
DNA
5′
5′
5′
5′
5′
5′
3′
3′
3′
3′
3′
3′
1
Sticky
end
GAATTC
CTTAAG
CTTAA
G AATTC
G
15. Figure 20.3-2
One possible combination
DNA fragment added
from another molecule
cut by same enzyme.
Base pairing occurs.
Restriction enzyme
cuts sugar-phosphate
backbones.
Restriction site
DNA
5′
5′
5′
5′
5′
5′
5′
5′
5′5′
5′
5′
5′5′
3′
3′
3′
3′
3′
3′
3′
3′
3′
3′
3′
3′
3′
3′
2
1
Sticky
end
GAATTC
CTTAAG
CTTAA
G AATTC
G
GG
AATTC
CTTAA
G
G
G
G
AATT CAATT C
C TTAA C TTAA
16. Figure 20.3-3
Recombinant DNA molecule
One possible combination
DNA ligase
seals strands
DNA fragment added
from another molecule
cut by same enzyme.
Base pairing occurs.
Restriction enzyme
cuts sugar-phosphate
backbones.
Restriction site
DNA
5′
5′
5′
5′
5′
5′
5′
5′
5′5′
5′
5′
5′5′
5′
5′
3′
3′
3′
3′
3′
3′
3′
3′
3′
3′
3′
3′
3′
3′
3′
3′
2
3
1
Sticky
end
GAATTC
CTTAAG
CTTAA
G AATTC
G
GG
AATTC
CTTAA
G
G
G
G
AATT CAATT C
C TTAA C TTAA
27. Figure 20.5
Foreign genome
Cut with restriction enzymes into either
small
fragments
large
fragments
or
Recombinant
plasmids
Plasmid
clone
(a) Plasmid library
(b) BAC clone
Bacterial artificial
chromosome (BAC)
Large
insert
with
many
genes
(c) Storing genome libraries
32. Figure 20.6-2
DNA in
nucleus
mRNAs in
cytoplasm
mRNA
Reverse
transcriptase Poly-A tail
DNA
strand
Primer
5′
5′
3′
3′
A A A A A A
T T T T T
33. Figure 20.6-3
DNA in
nucleus
mRNAs in
cytoplasm
mRNA
Reverse
transcriptase Poly-A tail
DNA
strand
Primer
5′
5′
5′
5′
3′
3′
3′
3′
A A A A A A
A A A A A A
T T T T T
T T T T T
34. Figure 20.6-4
DNA in
nucleus
mRNAs in
cytoplasm
mRNA
Reverse
transcriptase Poly-A tail
DNA
strand
Primer
DNA
polymerase
5′
5′
5′
5′
5′
5′
3′
3′
3′
3′
3′
3′
A A A A A A
A A A A A A
T T T T T
T T T T T
35. Figure 20.6-5
DNA in
nucleus
mRNAs in
cytoplasm
mRNA
Reverse
transcriptase Poly-A tail
DNA
strand
Primer
DNA
polymerase
cDNA
5′
5′
5′
5′
5′
5′
5′
5′
3′
3′
3′
3′
3′
3′
3′
3′
A A A A A A
A A A A A A
T T T T T
T T T T T
39. Figure 20.7
Radioactively
labeled probe
molecules Gene of
interest
Probe
DNA
Single-
stranded
DNA from
cell
Film
Location of
DNA with the
complementary
sequence
Nylon
membrane
Nylon membrane
Multiwell plates
holding library
clones
TECHNIQUE 5′
5′3′
3′
GAGTAGTGGCCG
⋅⋅⋅ CTCATCACCGGC⋅⋅⋅
54. Figure 20.9
Mixture of
DNA mol-
ecules of
different
sizes
Power
source
Power
source
Longer
molecules
Cathode Anode
Wells
Gel
Shorter
molecules
TECHNIQUE
RESULTS
1
2
− +
− +
55. Figure 20.9a
Mixture of
DNA mol-
ecules of
different
sizes
Power
source
Power
source
Longer
molecules
Cathode Anode
Wells
Gel
Shorter
molecules
TECHNIQUE
2
− +
− +
1
59. Figure 20.10
Normal β-globin allele
Sickle-cell mutant β-globin allele
Large
fragment
Normal
allele
Sickle-cell
allele
201 bp
175 bp
376 bp
(a) DdeI restriction sites in normal and
sickle-cell alleles of the β-globin gene
(b)Electrophoresis of restriction
fragments from normal and
sickle-cell alleles
201 bp175 bp
376 bp
Large fragment
Large fragment
DdeI DdeI DdeI DdeI
DdeI DdeI DdeI
60. Figure 20.10a
Normal β-globin allele
Sickle-cell mutant β-globin allele
(a) DdeI restriction sites in normal and
sickle-cell alleles of the β-globin gene
201 bp175 bp
376 bp
Large fragment
Large fragment
DdeI DdeI DdeI DdeI
DdeI DdeI DdeI
63. Figure 20.11
DNA + restriction enzyme
321
4
TECHNIQUE
I Normal
β-globin
allele
II Sickle-cell
allele
III Heterozygote
Restriction
fragments
Nitrocellulose
membrane (blot)
Heavy
weight
Gel
Sponge
Alkaline
solution Paper
towels
III III
III III III III
Preparation of
restriction fragments
Gel electrophoresis DNA transfer (blotting)
Radioactively labeled
probe for β-globin
gene
Nitrocellulose blot
Probe base-pairs
with fragments
Fragment from
sickle-cell
β-globin allele
Fragment from
normal β- globin
allele
Film
over
blot
Hybridization with labeled probe Probe detection5
65. Figure 20.12
DNA
(template strand)
TECHNIQUE
5′
3′
C
C
C
C
T
T
T
G
G
A
A
A
A
G
T
T
T
DNA
polymerase
Primer
5′
3′
P P P
OH
G
dATP
dCTP
dTTP
dGTP
Deoxyribonucleotides Dideoxyribonucleotides
(fluorescently tagged)
P P P
H
G
ddATP
ddCTP
ddTTP
ddGTP
5′
3′
C
C
C
C
T
T
T
G
G
A
A
A
A
DNA (template
strand)
Labeled strands
Shortest Longest
5′
3′
ddC
ddG
ddA
ddA
ddA
ddG
ddG
ddT
ddC
G
T
T
T
G
T
T
T
C
G
T
T
T
C
T T
G
G
T
T
T
C
T
G
A
G
T
T
T
C
T
G
A
A
G
T
T
T
C
T
G
A
A
G
G
T
T
T
C
T
G
A
A
G
T
G
T
T
T
C
T
G
A
A
G
T
C
G
T
T
T
C
T
G
A
A
G
T
C
A
Direction
of movement
of strands
Longest labeled strand
Detector
Laser
Shortest labeled strand
RESULTS
Last nucleotide
of longest
labeled strand
Last nucleotide
of shortest
labeled strand
G
G
G
A
A
A
C
C
T
66. Figure 20.12a
DNA
(template strand)
TECHNIQUE
Primer Deoxyribonucleotides Dideoxyribonucleotides
(fluorescently tagged)
DNA
polymerase
5′
5′
3′
3′
OH H
GG
dATP
dCTP
dTTP
dGTP
P P P P P P
ddATP
ddCTP
ddTTP
ddGTP
T
T
T
G
G
G
C
C
C
C
T
T
T
A
A
A
A
67. Figure 20.12b
DNA (template
strand)
Labeled strands
Shortest Longest
Direction
of movement
of strands
Longest labeled strand
Detector
Laser
Shortest labeled strand
TECHNIQUE (continued)
5′
3′
G
G
C
C
C
C
T
T
T
A
A
A
A
T
T
T
G
ddC
ddC
ddG
ddG
ddG
ddA
ddA
ddA
ddT
3′
5′
T
T
T
G
C
T
T
T
G
C
G
T
T
T
G
C
G
A
T
T
T
G
C
G
A
A
T
T
T
G
C
G
A
A
G
T
T
T
C
G
A
A
G
T
T
T
T
C
G
A
A
G
T
C
A
T
T
T
C
G
A
A
G
T
C
G G G
68. Figure 20.12c
RESULTS
Last nucleotide
of longest
labeled strand
Last nucleotide
of shortest
labeled strand
G
G
G
A
A
A
C
C
T
Direction
of movement
of strands
Longest labeled strand
Detector
Laser
Shortest labeled strand
77. Isolate mRNA.
2
1
3
4
TECHNIQUE
Make cDNA by reverse
transcription, using
fluorescently labeled
nucleotides.
Apply the cDNA mixture to a
microarray, a different gene
in each spot. The cDNA hybridizes
with any complementary DNA on
the microarray.
Rinse off excess cDNA; scan microarray
for fluorescence. Each fluorescent spot
(yellow) represents a gene expressed
in the tissue sample.
Tissue sample
mRNA molecules
Labeled cDNA molecules
(single strands)
DNA fragments
representing a
specific gene
DNA microarray
DNA microarray
with 2,400
human genes
Figure 20.15
85. Figure 20.17
Cross
section of
carrot root
2-mg
fragments
Fragments were
cultured in nu-
trient medium;
stirring caused
single cells to
shear off into
the liquid.
Single cells
free in
suspension
began to
divide.
Embryonic
plant developed
from a cultured
single cell.
Plantlet was
cultured on
agar medium.
Later it was
planted in soil.
Adult
plant
91. 4
5
6
RESULTS
Grown in culture
Implanted in uterus
of a third sheep
Embryonic
development
Nucleus from
mammary cell
Early embryo
Surrogate
mother
Lamb (“Dolly”) genetically
identical to mammary cell donor
Figure 20.19b
98. Figure 20.22
Remove skin cells
from patient. 2
1
3
4
Reprogram skin cells
so the cells become
induced pluripotent
stem (iPS) cells.
Patient with
damaged heart
tissue or other
disease
Return cells to
patient, where
they can repair
damaged tissue.
Treat iPS cells so
that they differentiate
into a specific
cell type.
103. Figure 20.23
Cloned gene
2
1
3
4
Retrovirus
capsid
Bone
marrow
cell from
patient
Viral RNA
Bone
marrow
Insert RNA version of normal allele
into retrovirus.
Let retrovirus infect bone marrow cells
that have been removed from the
patient and cultured.
Viral DNA carrying the normal
allele inserts into chromosome.
Inject engineered
cells into patient.
113. Figure 20.25
This photo shows
Washington just before
his release in 2001,
after 17 years in prison.
(a)
(b)These and other STR data exonerated Washington
and led Tinsley to plead guilty to the murder.
Semen on victim
Earl Washington
Kenneth Tinsley
17,19
16,18
17,19
13,16
14,15
13,16
12,12
11,12
12,12
Source of
sample
STR
marker 1
STR
marker 2
STR
marker 3
114. Figure 20.25a
This photo shows
Washington just before
his release in 2001,
after 17 years in prison.
(a)
118. Figure 20.26
Plant with new trait
RESULTS
TECHNIQUE
Ti
plasmid
Site where
restriction
enzyme cuts
DNA with
the gene
of interest
Recombinant
Ti plasmid
T DNA
Agrobacterium tumefaciens