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Molecular cloning takes advantage of the fact that the chemical structure of DNA is fundamentally the same in all living organisms. Therefore, if any segment of DNA from any organism is inserted into a DNA segment containing the molecular sequences required for DNA replication, and the resulting recombinant DNA is introduced into the organism from which the replication sequences were obtained ...
Golden Gate Cloning or Golden Gate assembly [1] 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. [2] This assembly is performed in vitro. Most commonly used Type IIS enzymes include BsaI, BsmBI, and ...
Protocols are presented step-by-step and edited in the style that has made Molecular Cloning, Antibodies, Cells and many other CSH manuals essential [tone] to the work of scientists worldwide. Protocols in the database come from CSH manuals, courses taught at Cold Spring Harbor Laboratory , and newly submitted methods from the scientific community.
Recombinant DNA (rDNA), or molecular cloning, is the process by which a single gene, or segment of DNA, is isolated and amplified. Recombinant DNA is also known as in vitro recombination . A cloning vector is a DNA molecule that carries foreign DNA into a host cell , where it replicates, producing many copies of itself along with the foreign DNA.
The choice of vector for molecular cloning depends on the choice of host organism, the size of the DNA to be cloned, and whether and how the foreign DNA is to be expressed. [7] The DNA segments can be combined by using a variety of methods, such as restriction enzyme/ligase cloning or Gibson assembly. [citation needed]
Gibson assembly is a molecular cloning method that allows for the joining of multiple DNA fragments in a single, isothermal reaction. It is named after its creator, Daniel G. Gibson, who is the chief technology officer and co-founder of the synthetic biology company, Telesis Bio.
Commonly made in the laboratory by solid-phase chemical synthesis, [1] these small fragments of nucleic acids can be manufactured as single-stranded molecules with any user-specified sequence, and so are vital for artificial gene synthesis, polymerase chain reaction (PCR), DNA sequencing, molecular cloning and as molecular probes.
Molecular Cloning Guides—References to help scientists design plasmid cloning experiments, including tutorials on restriction enzyme digestion and PCR-based cloning. Molecular Cloning Protocols—Specific protocols for a variety of plasmid cloning techniques, such as isolation of bacterial colonies, DNA purification by gel electrophoresis and ...
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