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Golden Gate Cloning
Golden Gate Cloning or Golden Gate assembly is a molecular cloning method that allows a researcher to simultaneously and directionally assemble multiple DNA fragments into a single piece using Type IIS restriction enzymes and T4 DNA ligase. This assembly is performed ''in vitro''. Most commonly used Type IIS enzymes include BsaI, BsmBI, and BbsI. Unlike standard Type II restriction enzymes like EcoRI and BamHI, these enzymes cut DNA outside of their recognition sites and, therefore, can create non-palindromic overhangs. Since 256 potential overhang sequences are possible, multiple fragments of DNA can be assembled by using combinations of overhang sequences. In practice, this means that Golden Gate Cloning is typically scarless. Additionally, because the final product does not have a Type IIS restriction enzyme recognition site, the correctly-ligated product cannot be cut again by the restriction enzyme, meaning the reaction is essentially irreversible. A typical thermal cyc ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Golden Gate Assembly
Golden Gate Cloning or Golden Gate assembly is a molecular cloning method that allows a researcher to simultaneously and directionally assemble multiple DNA fragments into a single piece using Type IIS restriction enzymes and T4 DNA ligase. This assembly is performed ''in vitro''. Most commonly used Type IIS enzymes include BsaI, BsmBI, and BbsI. Unlike standard Type II restriction enzymes like EcoRI and BamHI, these enzymes cut DNA outside of their recognition sites and, therefore, can create non-palindromic overhangs. Since 256 potential overhang sequences are possible, multiple fragments of DNA can be assembled by using combinations of overhang sequences. In practice, this means that Golden Gate Cloning is typically scarless. Additionally, because the final product does not have a Type IIS restriction enzyme recognition site, the correctly-ligated product cannot be cut again by the restriction enzyme, meaning the reaction is essentially irreversible. A typical thermal cyc ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Gateway Technology
Gateway cloning System, invented and commercialized by Invitrogen since the late 1990s, is a molecular biology method that enables researchers to efficiently transfer DNA-fragments between plasmids using a proprietary set of recombination sequences, the "Gateway att" sites, and two proprietary enzyme mixes, called "LR Clonase", and "BP Clonase". Gateway Cloning Technique allows transfer of DNA fragments between different cloning vectors while maintaining the reading frame. Using Gateway, one can clone subclone DNA segments for functional analysis. The system requires the initial insertion of a DNA fragment into a plasmid with two flanking recombination sequences called “att L 1” and “att L 2”, to develop a “Gateway Entry clone” (special Invitrogen nomenclature). Large archives of Gateway Entry clones, containing the vast majority of human, mouse and rat ORFs (open reading frames) have been cloned from human cDNA libraries or chemically synthesized to support the resear ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Golden Gate Bridge
The Golden Gate Bridge is a suspension bridge spanning the Golden Gate, the strait connecting San Francisco Bay and the Pacific Ocean. The structure links the U.S. city of San Francisco, California—the northern tip of the San Francisco Peninsula—to Marin County, carrying both U.S. Route 101 and California State Route 1 across the strait. It also carries pedestrian and bicycle traffic, and is designated as part of U.S. Bicycle Route 95. Being declared one of the Wonders of the Modern World by the American Society of Civil Engineers, the bridge is one of the most internationally recognized symbols of San Francisco and California. It was initially designed by engineer Joseph Strauss in 1917. The bridge was named for the Golden Gate strait, the channel that it spans. The Frommer's travel guide describes the Golden Gate Bridge as "possibly the most beautiful, certainly the most photographed, bridge in the world." At the time of its opening in 1937, it was both the longe ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Gateway Technology
Gateway cloning System, invented and commercialized by Invitrogen since the late 1990s, is a molecular biology method that enables researchers to efficiently transfer DNA-fragments between plasmids using a proprietary set of recombination sequences, the "Gateway att" sites, and two proprietary enzyme mixes, called "LR Clonase", and "BP Clonase". Gateway Cloning Technique allows transfer of DNA fragments between different cloning vectors while maintaining the reading frame. Using Gateway, one can clone subclone DNA segments for functional analysis. The system requires the initial insertion of a DNA fragment into a plasmid with two flanking recombination sequences called “att L 1” and “att L 2”, to develop a “Gateway Entry clone” (special Invitrogen nomenclature). Large archives of Gateway Entry clones, containing the vast majority of human, mouse and rat ORFs (open reading frames) have been cloned from human cDNA libraries or chemically synthesized to support the resear ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Yuri Gleba
Yuri may refer to: People and fictional characters Given name *Yuri (Slavic name), the Slavic masculine form of the given name George, including a list of people with the given name Yuri, Yury, etc. * Yuri (Japanese name), also Yūri, feminine Japanese given names, including a list of people and fictional characters *Yu-ri (Korean name), Korean unisex given name, including a list of people and fictional characters Singers *Yuri (Japanese singer), vocalist of the band Move *Yuri (Korean singer), member of Girl Friends * Yuri (Mexican singer) *Kwon Yu-ri, member of Girls' Generation Footballers *Yuri (footballer, born 1982), full name Yuri de Souza Fonseca, Brazilian football forward *Yuri (footballer, born 1984), full name Yuri Adriano Santos, Brazilian footballer *Yuri (footballer, born 1986), full name Yuri Vera Cruz Erbas, Brazilian footballer *Yuri (footballer, born 1989), full name Yuri Naves Roberto, Brazilian football defensive midfielder *Yuri (footballer, born 1990), full ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Agrobacterium
''Agrobacterium'' is a genus of Gram-negative bacteria established by H. J. Conn that uses horizontal gene transfer to cause tumors in plants. ''Agrobacterium tumefaciens'' is the most commonly studied species in this genus. ''Agrobacterium'' is well known for its ability to transfer DNA between itself and plants, and for this reason it has become an important tool for genetic engineering. Nomenclatural History Leading up to the 1990s, the genus ''Agrobacterium'' was used as a wastebasket taxon. With the advent of 16S sequencing, many ''Agrobacterium'' species (especially the marine species) were reassigned to genera such as ''Ahrensia'', ''Pseudorhodobacter'', ''Ruegeria'', and ''Stappia''. The remaining ''Agrobacterium'' species were assigned to three biovars: biovar 1 (''Agrobacterium tumefaciens''), biovar 2 (''Agrobacterium rhizogenes''), and biovar 3 (''Agrobacterium vitis''). In the early 2000s, ''Agrobacterium'' was synonymized with the genus ''Rhizobium''. This move pr ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Schematic Workflow For Generating Complex Combinatorial DNA Libraries
A schematic, or schematic diagram, is a designed representation of the elements of a system using abstract, graphic symbols rather than realistic pictures. A schematic usually omits all details that are not relevant to the key information the schematic is intended to convey, and may include oversimplified elements in order to make this essential meaning easier to grasp, as well as additional organization of the information. For example, a subway map intended for passengers may represent a subway station with a dot. The dot is not intended to resemble the actual station at all but aims to give the viewer information without unnecessary visual clutter. A schematic diagram of a chemical process uses symbols in place of detailed representations of the vessels, piping, valves, pumps, and other equipment that compose the system, thus emphasizing the functions of the individual elements and the interconnections among them and suppresses their physical details. In an electronic circuit ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Protein Engineering
Protein engineering is the process of developing useful or valuable proteins. It is a young discipline, with much research taking place into the understanding of protein folding and recognition for protein design principles. It has been used to improve the function of many enzymes for industrial catalysis. It is also a product and services market, with an estimated value of $168 billion by 2017. There are two general strategies for protein engineering: rational protein design and directed evolution. These methods are not mutually exclusive; researchers will often apply both. In the future, more detailed knowledge of protein structure and function, and advances in high-throughput screening, may greatly expand the abilities of protein engineering. Eventually, even unnatural amino acids may be included, via newer methods, such as expanded genetic code, that allow encoding novel amino acids in genetic code. Approaches Rational design In rational protein design, a scientist uses ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
BioBrick
BioBrick parts are DNA sequences which conform to a restriction-enzyme assembly standard. These building blocks are used to design and assemble larger synthetic biological circuits from individual parts and combinations of parts with defined functions, which would then be incorporated into living cells such as ''Escherichia coli'' cells to construct new biological systems. Examples of BioBrick parts include promoters, ribosomal binding sites (RBS), coding sequences and terminators. Overview The BioBrick parts are used by applying engineering principles of abstraction and modularization. BioBrick parts form the base of the hierarchical system on which synthetic biology is based. There are three levels to the hierarchy: # Parts: Pieces of DNA that form a functional unit (for example promoter, RBS, etc.) # Device: Collection set of parts with defined function. In simple terms, a set of complementary BioBrick parts put together forms a device. # System: Combination of a set of d ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Methods In Molecular Biology
''Methods in Molecular Biology'' is a book series published by Humana Press (an imprint of Springer Science+Business Media) that covers molecular biology research methods and protocols. The book series was introduced by series editor John M. Walker in 1983 and provides step-by-step instructions for carrying out experiments in a research lab. As of January 2020, more than 2000 volumes (2471 as of 10-March-2022) had been published in the series. The protocols are also available online in SpringerLink, and were previously in ''Springer Protocols ''Springer Protocols'' was a database of life sciences protocols published by Springer Science+Business Media. It replaced ''BioMed Protocols'', a Humana Press database, in January 2008, and was deactivated on 25 July 2018. The protocols were then ...''. Each protocol opens with an introductory overview and a list of the materials and reagents needed to complete the experiment. Every protocol is followed by a detailed procedure that is supp ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Molecular Cloning
Molecular cloning is a set of experimental methods in molecular biology that are used to assemble recombinant DNA molecules and to direct their replication within host organisms. The use of the word ''cloning'' refers to the fact that the method involves the replication of one molecule to produce a population of cells with identical DNA molecules. Molecular cloning generally uses DNA sequences from two different organisms: the species that is the source of the DNA to be cloned, and the species that will serve as the living host for replication of the recombinant DNA. Molecular cloning methods are central to many contemporary areas of modern biology and medicine. In a conventional molecular cloning experiment, the DNA to be cloned is obtained from an organism of interest, then treated with enzymes in the test tube to generate smaller DNA fragments. Subsequently, these fragments are then combined with vector DNA to generate recombinant DNA molecules. The recombinant DNA is then in ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Thermal Cycler
The thermal cycler (also known as a thermocycler, PCR machine or DNA amplifier) is a laboratory apparatus most commonly used to amplify segments of DNA via the polymerase chain reaction (PCR). Thermal cyclers may also be used in laboratories to facilitate other temperature-sensitive reactions, including restriction enzyme digestion or rapid diagnostics. The device has a ''thermal block'' with holes where tubes holding the reaction mixtures can be inserted. The cycler then raises and lowers the temperature of the block in discrete, pre-programmed steps. History The earliest thermal cyclers were designed for use with the Klenow fragment of DNA polymerase I. Since this enzyme is destroyed during each heating step of the amplification process, new enzyme had to be added every cycle. This led to a cumbersome machine based on an automated pipettor, with open reaction tubes. Later, the PCR process was adapted to the use of thermostable DNA polymerase from ''Thermus aquaticus'', w ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
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