Cloning plasmid
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This document describes plasmids as cloning vectors. It outlines that plasmids are small, circular, self-replicating DNA molecules found in prokaryotes. Important features of plasmids for use as cloning vectors include a cloning site, origin of replication, and selectable marker like antibiotic resistance. Plasmids are useful for cloning small DNA fragments but less so for larger fragments. They provide advantages like easy selection strategies but a disadvantage is less usefulness for large DNA cloning.
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Molecular cloning vectors
The molecular cloning vectors used in genetic engineering-plasmids,phage vectors,Hbrid vectors and artificial chromosomes
Bacteriophage vectors
This document describes the bacteriophages λ and M13, which are commonly used as cloning vectors. λ phage is a temperate phage that infects E. coli and has a double-stranded linear DNA genome. Its genome is organized into regions that encode proteins for the phage head, tail, and lysogeny/lysis functions. M13 is a filamentous phage with a single-stranded circular genome. Both phages can be modified to serve as insertion or replacement vectors for cloning foreign DNA fragments into E. coli.
Restriction enzyme
Enzymes that cut DNA at or near specific recognition nucleotide sequences known as restriction sites.
Especial class of enzymes that cleave (cut) DNA at a specific unique internal location along its length.
Often called restriction endonucleases (Because they cut within the molecule).
Discovered in the late 1970s by Werner Arber, Hamilton Smith, and Daniel Nathans.
Essential tools for recombinant DNA technology.
Naturally produced by bacteria that use them as a defense mechanism against viral infection.
Chop up the viral nucleic acids and protect a bacterial cell by hydrolyzing phage DNA.
Cloning vectors ppt kiran b k final
Cloning vectors are small DNA molecules used to replicate, amplify and express inserted DNA fragments. There are several types of cloning vectors including plasmids, bacteriophages, cosmids, and artificial chromosomes. Plasmids are the most commonly used cloning vectors as they can replicate autonomously in bacterial cells, contain selectable markers, and accept DNA insert sizes up to 10kb. Bacteriophages such as lambda can accept larger inserts up to 20kb but have a narrow host range. Cosmids combine properties of plasmids and phages to accept inserts up to 50kb.
Chromosome walking
Chromosome walking is a method used to isolate and clone a particular gene or allele through positional cloning. It involves using overlapping clones that contain DNA fragments near the target gene to "walk" through the chromosome until reaching the gene. Each successive clone is tested to map its precise location until eventually reaching the target gene. Chromosome walking was developed in the early 1980s and can be used to analyze genetically transmitted diseases and find single nucleotide polymorphisms. However, it has limitations such as being a slow process and difficulty walking through repeated sequences.
Vector construction
Vectors are DNA molecules that can accept foreign DNA and be replicated within a host cell. They are required for cloning genes and transferring them to bacteria. Common vectors include plasmids, bacteriophages, cosmids, and artificial chromosomes. Expression vectors are used to produce proteins from cloned DNA, and come in prokaryotic and eukaryotic varieties. Cloning vectors replicate recombinant DNA within host cells to produce multiple clones and can accommodate different sized DNA fragments depending on the vector type.
Molecular Cloning - Vectors: Types & Characteristics
Vectors are DNA molecules that can carry foreign DNA fragments into host cells. There are two main types of vectors: cloning vectors for propagating DNA inserts and expression vectors for expressing inserted DNA. Common vector types include plasmids, bacteriophages, cosmids, and artificial chromosomes. Vectors must have key characteristics like the ability to self-replicate, selectable markers, and origins of replication to successfully clone and transfer DNA.
Plasmid as a Cloning Vector
Plasmids are circular DNA molecules that can exist independently of the bacterial chromosome and are used as cloning vectors. Cloning vectors are DNA molecules that can carry foreign genetic material into a host cell. Plasmids have features like cloning sites, selectable markers, and reporter genes that make them useful for cloning. Common types of cloning vectors include plasmids, bacteriophages, cosmids, and artificial chromosomes. Plasmids are advantageous as cloning vectors because they are small, can replicate independently of the host, and often confer antibiotic resistance, allowing for selection of transformed cells.
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Molecular cloning vectors
The molecular cloning vectors used in genetic engineering-plasmids,phage vectors,Hbrid vectors and artificial chromosomes
Bacteriophage vectors
This document describes the bacteriophages λ and M13, which are commonly used as cloning vectors. λ phage is a temperate phage that infects E. coli and has a double-stranded linear DNA genome. Its genome is organized into regions that encode proteins for the phage head, tail, and lysogeny/lysis functions. M13 is a filamentous phage with a single-stranded circular genome. Both phages can be modified to serve as insertion or replacement vectors for cloning foreign DNA fragments into E. coli.
Restriction enzyme
Enzymes that cut DNA at or near specific recognition nucleotide sequences known as restriction sites.
Especial class of enzymes that cleave (cut) DNA at a specific unique internal location along its length.
Often called restriction endonucleases (Because they cut within the molecule).
Discovered in the late 1970s by Werner Arber, Hamilton Smith, and Daniel Nathans.
Essential tools for recombinant DNA technology.
Naturally produced by bacteria that use them as a defense mechanism against viral infection.
Chop up the viral nucleic acids and protect a bacterial cell by hydrolyzing phage DNA.
Cloning vectors ppt kiran b k final
Cloning vectors are small DNA molecules used to replicate, amplify and express inserted DNA fragments. There are several types of cloning vectors including plasmids, bacteriophages, cosmids, and artificial chromosomes. Plasmids are the most commonly used cloning vectors as they can replicate autonomously in bacterial cells, contain selectable markers, and accept DNA insert sizes up to 10kb. Bacteriophages such as lambda can accept larger inserts up to 20kb but have a narrow host range. Cosmids combine properties of plasmids and phages to accept inserts up to 50kb.
Chromosome walking
Chromosome walking is a method used to isolate and clone a particular gene or allele through positional cloning. It involves using overlapping clones that contain DNA fragments near the target gene to "walk" through the chromosome until reaching the gene. Each successive clone is tested to map its precise location until eventually reaching the target gene. Chromosome walking was developed in the early 1980s and can be used to analyze genetically transmitted diseases and find single nucleotide polymorphisms. However, it has limitations such as being a slow process and difficulty walking through repeated sequences.
Vector construction
Vectors are DNA molecules that can accept foreign DNA and be replicated within a host cell. They are required for cloning genes and transferring them to bacteria. Common vectors include plasmids, bacteriophages, cosmids, and artificial chromosomes. Expression vectors are used to produce proteins from cloned DNA, and come in prokaryotic and eukaryotic varieties. Cloning vectors replicate recombinant DNA within host cells to produce multiple clones and can accommodate different sized DNA fragments depending on the vector type.
Molecular Cloning - Vectors: Types & Characteristics
Vectors are DNA molecules that can carry foreign DNA fragments into host cells. There are two main types of vectors: cloning vectors for propagating DNA inserts and expression vectors for expressing inserted DNA. Common vector types include plasmids, bacteriophages, cosmids, and artificial chromosomes. Vectors must have key characteristics like the ability to self-replicate, selectable markers, and origins of replication to successfully clone and transfer DNA.
Plasmid as a Cloning Vector
Plasmids are circular DNA molecules that can exist independently of the bacterial chromosome and are used as cloning vectors. Cloning vectors are DNA molecules that can carry foreign genetic material into a host cell. Plasmids have features like cloning sites, selectable markers, and reporter genes that make them useful for cloning. Common types of cloning vectors include plasmids, bacteriophages, cosmids, and artificial chromosomes. Plasmids are advantageous as cloning vectors because they are small, can replicate independently of the host, and often confer antibiotic resistance, allowing for selection of transformed cells.
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PHAGEMID.pdf
Phagemids are cloning vectors that have properties of both plasmids and bacteriophages. They contain an origin of replication for double-stranded DNA replication as plasmids do, as well as an f1 origin that allows for single-stranded replication and packaging into phage particles. Phagemids are used for cloning, expression, sequencing, and generating RNA probes. They offer advantages over other phage vectors such as larger cloning capacity and stability. A typical phagemid contains origins of replication from phage M13 and a plasmid, antibiotic resistance, a multiple cloning site, and phage promoters.
Restriction enzymes genetic enginering
Restriction enzymes are molecular scissors that cut DNA at specific recognition sequences. They are found in bacteria and archaea and serve to defend the host genome from foreign DNA like viruses. There are four main types of restriction enzymes that differ in composition, cofactor requirements, target sequence, and cleavage site. Restriction enzymes are important tools in biotechnology for manipulating DNA through cutting and joining of fragments. They allow for processes like DNA cloning, mapping, and sequencing.
lambda cloning vector
- Lambda bacteriophage cloning vectors were developed to overcome limitations in the size of DNA that could be inserted into unmodified lambda vectors.
- Segments of the non-essential lambda genome could be deleted to allow insertion of up to 18kb of new DNA while still allowing packaging.
- Natural selection was used to generate lambda strains lacking restriction sites, allowing restriction-based cloning.
- The first lambda vectors were insertion and replacement vectors, while later cosmids allowed cloning of fragments up to 52kb.
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The document discusses transgenic mice and fish. It defines transgenic animals as those permanently engineered through gene insertion. Methods to create transgenic mice include retroviral vectors, DNA microinjection into pronuclei, and using engineered embryonic stem cells. Transgenic mice have aided research in many areas. Transgenic fish are also discussed, including important salmon and tilapia examples. Gene flow and food safety are concerns for transgenic animals.
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3. The history of antisense technology was discussed, including its first observation in nature's HOK/SOK system and early experiments in the 1990s that helped define gene silencing.
Cosmids vector
1. A cosmid is a type of plasmid vector that contains sequences from bacteriophage lambda, specifically the cos sites. This allows DNA fragments up to 45kb to be packaged into phage particles and transduced into bacteria.
2. Cosmids are developed by combining features of plasmid and phage vectors. They can accept large DNA inserts through packaging and transduction while also replicating stably inside bacteria like plasmids.
3. Cloning with cosmids involves inserting DNA fragments into the cosmid, ligating to form concatemers, in vitro packaging into phage particles, and transducing the particles into bacteria where the cosmids replicate as plasmids.
Bioinformatics
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The other vectors which are generally used are DNA and RNA viruses.
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The document discusses various "omics" fields of study including genomics, proteomics, metabolomics, and others. It provides definitions and descriptions of each type of omics, focusing on the large sets of biological molecules they each study such as genomes, proteomes, metabolomes, etc. It explains that omics fields examine biological data on a large scale and provide insights into biological processes, functions, and interactions on a systems-wide level.
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Retroviral Vectors
Retroviral vectors are derived from wild type retroviruses like Moloney murine leukemia virus. They are engineered to carry foreign genes into target cells. The vectors contain cis-acting elements from the viral genome like LTRs and packaging signals but lack the trans-acting genes gag, pol, and env. This prevents replication but allows the vector and its gene of interest to integrate stably into the host cell genome. Retroviral vectors show promise for gene therapy applications but also have limitations like requiring actively dividing cells for transduction and potential risks of insertional mutagenesis leading to cancer.
Gene cloning presentation
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Histotypic culture
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Cloning Vectors (plasmid).pdf
Plasmids are circular, self-replicating DNA molecules that are commonly used as cloning vectors. Key properties of plasmids that make them useful for cloning include their ability to replicate independently of the bacterial chromosome, carry genes for antibiotic resistance or other selectable markers, and accept foreign DNA at specific restriction sites. Common plasmids used for cloning are pBR322, which has features like a high copy number and blue-white screening, and pUC plasmids, which have even higher copy numbers. Plasmids can shuttle DNA between bacterial and yeast cells, and RNA production plasmids contain promoters to transcribe cloned DNA into RNA for applications like Northern blotting.
Cloning by bacterial plasmids
Plasmid vectors are circular, self-replicating DNA molecules found in bacteria cytoplasm that can transfer genes between cells. They are useful for cloning small DNA fragments under 10kb due to their small size and selection strategies. However, plasmids are less useful for cloning large fragments over 10kb. Plasmid vectors for cloning contain an origin of replication for independent replication, selective markers for identifying cells containing the plasmid, and a multiple cloning site to insert DNA fragments.
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This document discusses the patentability of live organisms. It begins by explaining what patents are and the criteria for patentability. It then discusses how traditionally only inventions could be patented, but now living things like cells, bacteria, and extracts from plants have also been patented. The US initially only allowed patents on microorganisms, but later expanded this to include animals and modified life forms. Several controversial patents have been granted, such as on the OncoMouse and human genes. While many see issues with patenting life, proponents argue it encourages innovation. The document reviews patenting scenarios in countries like the US, India, and under international agreements.
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The document summarizes phagemid and bacterial artificial chromosome (BAC) vectors. It describes that phagemid vectors are plasmids that contain both plasmid and phage origins of replication. Specifically, it discusses the features of pBluescript II phagemid vectors, including their polylinker and RNA polymerase promoter sequences. It also describes how pBluescript II phagemid vectors can produce blue or white colonies depending on insert presence. The document then explains that BAC vectors are low-copy plasmids that can hold up to 300kb DNA fragments. Examples of BAC vectors like pBAC108L and pBeloBAC11 are provided, with details about their replication origin and partitioning functions.
PHAGEMID.pdf
Phagemids are cloning vectors that have properties of both plasmids and bacteriophages. They contain an origin of replication for double-stranded DNA replication as plasmids do, as well as an f1 origin that allows for single-stranded replication and packaging into phage particles. Phagemids are used for cloning, expression, sequencing, and generating RNA probes. They offer advantages over other phage vectors such as larger cloning capacity and stability. A typical phagemid contains origins of replication from phage M13 and a plasmid, antibiotic resistance, a multiple cloning site, and phage promoters.
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Restriction enzymes are molecular scissors that cut DNA at specific recognition sequences. They are found in bacteria and archaea and serve to defend the host genome from foreign DNA like viruses. There are four main types of restriction enzymes that differ in composition, cofactor requirements, target sequence, and cleavage site. Restriction enzymes are important tools in biotechnology for manipulating DNA through cutting and joining of fragments. They allow for processes like DNA cloning, mapping, and sequencing.
lambda cloning vector
- Lambda bacteriophage cloning vectors were developed to overcome limitations in the size of DNA that could be inserted into unmodified lambda vectors.
- Segments of the non-essential lambda genome could be deleted to allow insertion of up to 18kb of new DNA while still allowing packaging.
- Natural selection was used to generate lambda strains lacking restriction sites, allowing restriction-based cloning.
- The first lambda vectors were insertion and replacement vectors, while later cosmids allowed cloning of fragments up to 52kb.
Transgenic animals, mice and fish
The document discusses transgenic mice and fish. It defines transgenic animals as those permanently engineered through gene insertion. Methods to create transgenic mice include retroviral vectors, DNA microinjection into pronuclei, and using engineered embryonic stem cells. Transgenic mice have aided research in many areas. Transgenic fish are also discussed, including important salmon and tilapia examples. Gene flow and food safety are concerns for transgenic animals.
Transgenic plants
This document summarizes an assignment on applications of transgenic plants. It first introduces transgenic plant technology and how genes can be artificially inserted into plant genomes. It then lists several applications of transgenic plants, including herbicide and insect resistance, virus resistance, altered oil content, and delayed fruit ripening. Specific examples are provided, such as Bt cotton providing insect resistance and Roundup Ready soybeans providing herbicide tolerance. The document also discusses transgenic plants for fungal/bacterial resistance using PR proteins, as well as virus resistant plants and methods for controlling plant pollination.
Antisense RNA in crop
This document summarizes a seminar presentation on antisense RNA technology. The presentation covered:
1. The introduction defined antisense RNA and its potential for crop improvement to meet rising global food demand.
2. The mechanisms of antisense RNA technology were explained, including how antisense RNA binds to mRNA to inhibit translation and activate RNase H degradation.
3. The history of antisense technology was discussed, including its first observation in nature's HOK/SOK system and early experiments in the 1990s that helped define gene silencing.
Cosmids vector
1. A cosmid is a type of plasmid vector that contains sequences from bacteriophage lambda, specifically the cos sites. This allows DNA fragments up to 45kb to be packaged into phage particles and transduced into bacteria.
2. Cosmids are developed by combining features of plasmid and phage vectors. They can accept large DNA inserts through packaging and transduction while also replicating stably inside bacteria like plasmids.
3. Cloning with cosmids involves inserting DNA fragments into the cosmid, ligating to form concatemers, in vitro packaging into phage particles, and transducing the particles into bacteria where the cosmids replicate as plasmids.
Bioinformatics
This document discusses gene prediction and promoter prediction. It begins by explaining that gene prediction involves locating protein-coding genes within sequenced genomes in order to understand their functional content. Various computational methods are used for gene prediction, including searching for signals like start/stop codons, searching coding content, and comparing sequences to find homologs. Promoter prediction involves locating DNA elements that regulate gene expression and is challenging due to diversity and short, conserved motifs. Ab initio and comparative phylogenetic footprinting methods are used to predict promoters and regulatory elements in prokaryotes and eukaryotes.
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What are an expression vector? Detailed description of plant gene structure. Plant expression vector systems are generally consists of Ri and Ti plasmids.
The other vectors which are generally used are DNA and RNA viruses.
Omics era
The document discusses various "omics" fields of study including genomics, proteomics, metabolomics, and others. It provides definitions and descriptions of each type of omics, focusing on the large sets of biological molecules they each study such as genomes, proteomes, metabolomes, etc. It explains that omics fields examine biological data on a large scale and provide insights into biological processes, functions, and interactions on a systems-wide level.
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properties,
quality of good vectors,
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Differential gene profiling methods
This document discusses differential gene profiling methods, specifically differential display and subtractive hybridization. It provides details on how differential display and subtractive hybridization work, including that differential display allows comparison of gene expression between two samples and was commonly used in the 1990s before being replaced by microarrays in 2000. Subtractive hybridization identifies differences in nucleic acids between two populations and enriches for sequences not common to both. The document discusses applications and references for further information.
Retroviral Vectors
Retroviral vectors are derived from wild type retroviruses like Moloney murine leukemia virus. They are engineered to carry foreign genes into target cells. The vectors contain cis-acting elements from the viral genome like LTRs and packaging signals but lack the trans-acting genes gag, pol, and env. This prevents replication but allows the vector and its gene of interest to integrate stably into the host cell genome. Retroviral vectors show promise for gene therapy applications but also have limitations like requiring actively dividing cells for transduction and potential risks of insertional mutagenesis leading to cancer.
Gene cloning presentation
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Histotypic culture
Histotypic culture involves growing cell lines in a three-dimensional matrix at high density to form tissue-like structures. Common techniques include using gels, sponges, hollow fibers, spheroids, and rotating chambers to provide a 3D environment that allows cells to organize similarly to how they would in tissues. This allows for the study of processes like drug penetration and cell differentiation that are not possible with traditional 2D cultures. While histotypic cultures provide a model for certain tissue functions, they also face challenges like loss of cell differentiation over time.
Genomic library
Genomic libraries are collections of bacteria that have been engineered to contain the entire DNA of an organism. DNA from the organism is cut into fragments and inserted into cloning vectors like plasmids within bacteria. Together the bacteria hold the complete genome. Genomic libraries allow researchers to isolate and study specific DNA sequences using probes, map genomes, and clone DNA segments for various research purposes like genetic modification of crops. They are constructed by extracting, cutting, and ligating an organism's DNA into vectors within host bacteria. Different vectors have varying capacities to hold DNA fragments, ranging from plasmids at 15kb to yeast artificial chromosomes at up to 2,000kb.
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Plasmid vectors are circular, self-replicating DNA molecules found in bacteria cytoplasm that can transfer genes between cells. They are useful for cloning small DNA fragments under 10kb due to their small size and selection strategies. However, plasmids are less useful for cloning large fragments over 10kb. Plasmid vectors for cloning contain an origin of replication for independent replication, selective markers for identifying cells containing the plasmid, and a multiple cloning site to insert DNA fragments.
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The document discusses various types of plasmid vectors and cloning vectors used for gene cloning. It describes the key properties required for a vector, including autonomous replication, small size, selectable markers, and restriction enzyme sites. Some examples of early plasmid vectors discussed are pSC101, ColE1, and pBR322. Later vectors with improved properties include the pUC series, pGEM series, and pET series. A variety of vectors were also developed using bacteriophages, cosmids, artificial chromosomes, and more to clone larger fragments.
Lectut btn-202-ppt-l3. gene cloning and plasmid vectors (1)
The document discusses various types of plasmid vectors and cloning vectors used for gene cloning. It describes the key properties required for a vector, including autonomous replication, small size, selectable markers, and restriction enzyme sites. Some examples of early plasmid vectors discussed are pSC101, ColE1, and pBR322. Later vectors with improved properties include the pUC series, pGEM series, and pET series. A variety of other vector types were also constructed for different applications, such as bacteriophages, cosmids, YACs, BACs, and artificial chromosomes.
Cloning vector
This document discusses cloning vectors, which are DNA molecules used to transport cloned DNA sequences between biological hosts. It defines a cloning vector as a small piece of DNA from a virus, plasmid, or cell that can maintain foreign DNA for cloning. The summary describes the key features of cloning vectors, including an origin of replication, cloning site, selectable marker, and optional reporter gene. It also lists common vector types like plasmids, bacteriophages, cosmids, and artificial chromosomes, and factors that determine the choice of vector, such as insert size.
Cloning vectors
Objectives:
After the end of the presentation we’ll know -
What is cloning vector?
Why cloning vector?
History
Features of a cloning vector
Types of cloning vector
Plasmid
Bacteriophage
Cosmid
Bacterial Artificial Chromosome (BAC)
Yeast Artificial Chromosome (BAC)
Human Artificial Chromosome (HAC)
Retroviral Vectors
What determines choice of vector?
Vector in molecular gene cloning
Cloning vector - The molecular analysis of DNA has been made possible by the cloning of DNA. The two molecules that are required for cloning are the DNA to be cloned and a cloning vector.
A cloning vector is a small piece of DNA taken from a virus, a plasmid or the cell of a higher organism, that can be stably maintained in an organism and into which a foreign DNA fragment can be inserted for cloning purposes.
Most vectors are genetically engineered.
The cloning vector is chosen according to the size and type of DNA to be cloned.
The vector therefore contains features that allow for the convenient insertion or removal of DNA fragment in or out of the vector, for example by treating the vector and the foreign DNA with a restriction enzyme and then ligating the fragments together.
After a DNA fragment has been cloned into a cloning vector, it may be further subcloned into another vector designed for more specific use.
Cloning vector series 1
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
Lecture_3-Vectors.pdf
The document discusses cloning vectors. It describes what a cloning vector is, including that it is a small piece of DNA that can stably maintain foreign DNA for cloning purposes. Common types of cloning vectors are described in detail, including plasmids, bacteriophages, cosmids, yeast artificial chromosomes, bacterial artificial chromosomes, and plant virus vectors. Key features of cloning vectors like origins of replication, antibiotic resistance genes, and cloning sites are also summarized.
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Cloning plasmid
- 1. PLASMID AS CLONING VECTOR Jean de Dieu BAZIKI PhD Scholar
- 2. OUTLINE • Introduction • Features • Types • Advantages • Disadvantages • References
- 3. Objective Describe salient features for ideal plasmid as cloning vector
- 4. Introduction Plasmid Extrachromosal Self replicating Short double stranded Closed Circular DNA molecules Found in prokaryotes Small size
- 5. Types of plasmids by their Functions Col plasmids Fertility-F plasmids Resistance plasmids Degradative plasmids Virulence plasmids Others Ti plasmids Ri plasmids
- 6. Plasmids as cloning vectors In 1973, Chang and Cohen proved the use of plasmid as cloning vector in genetic engineering/ Biotechnology .
- 7. Ideal features plasmid as cloning vector Small size 1.2-3kb Origin of replication Unique restriction site Multiple cloning site Selection marker Copy number Control elements ( Promoter, operator, ribosomal binding sites) No pathogenicity
- 10. 3 important features: Cloning site, Ori-an origin of replication, A selectable marker (ampr)
- 13. Advantages • Small, easy to handle • Straightforward selection strategies • Useful for cloning small DNA fragments(< 10kbp)
- 14. Disadvantages • Less useful for cloning large DNA fragments (> 10kbp)
- 15. Application A particular gene can be isolated and its nucleotide sequence be determined Genomic library construction Preparation of probes Control sequences DNA can be identified and analysed Protein/enzyme/RNA function can be investigated Mutations can be identified e.g: gene defects related to specific diseases Organism can be “engineered” for specific purposes e.g: Insulin production
- 16. References Cohen, S. N., Chang, A. C., Boyer, H. W., & Helling, R. B. (1973). Construction of biologically functional bacterial plasmids in vitro. Proceedings of the National Academy of Sciences of the United States of America, 70(11), 3240–4. Retrieved from http://www.ncbi.nlm.nih.gov/pubmed/4594039 full-text. (n.d.). Gene Cloning and DNA Analysis: An Introduction - T. A. Brown - Google Books. (n.d.). Retrieved February 24, 2017, from https://books.google.co.ke/books?hl=en&lr=&id=pi3dCQAAQBAJ&oi=fnd&pg=PR17 &dq=plasmid+cloning+vector+features&ots=C3_dR165S5&sig=LM0J_IErGQcSujVDa KydASzErg8&redir_esc=y#v=onepage&q=plasmid cloning vector features&f=false Promega. (2010). pGEM®- T and pGEM®- T Easy Vector Systems. Technical Manual, 1– 28.
- 17. THANK YOU MERCI MURAKOZE (Rwanda) ASANTE SANA (Kenya) NAMASIGNALO (Ethiopia) YEKENIYELLEY (Eritrea) GANGI DEBE (Chad) MASIAKNEE (Cameroon) WEBALE (Uganda)