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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 ...
Description: en: Reproductive and therapeutic cloning english diagram / de: Reproduktives und therapeutisches Klonen mit englicsh Text / Somatic body cell with desired genes, Nucleus fused with enucleated egg cell, Clone, Egg cell, Nucleus removed, REPRODUCTIVE CLONING, THERAPEUTIC CLONING, Surrogate mother, Tissue culture
An alternative technique called "handmade cloning" was described by Indian scientists in 2001. This technique requires no use of a micromanipulator and has been used for the cloning of several livestock species. [11] Removal of the nucleus can be done chemically, by centrifuge, or with the use of a blade.
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.
Nuclear transfer is a delicate process that is a major hurdle in the development of cloning technology. [5] Materials used in this procedure are a microscope, a holding pipette (small vacuum) to keep the oocyte in place, and a micropipette (hair-thin needle) capable of extracting the nucleus of a cell using a vacuum.
Usually the ultimate aim of expression cloning is to produce large quantities of specific proteins.To this end, a bacterial expression clone may include a ribosome binding site (Shine-Dalgarno sequence) to enhance translation of the gene of interest's mRNA, a transcription termination sequence, or, in eukaryotes, specific sequences to promote the post-translational modification of the protein ...
Use the enzyme DNA ligase to seal the DNA fragments into the vector. This creates a large pool of recombinant molecules. These recombinant molecules are taken up by a host bacterium by transformation, creating a DNA library. [9] [10] Below is a diagram of the above outlined steps. Genomic Library Construction
Cloning is commonly used to amplify DNA fragments containing whole genes, but it can also be used to amplify any DNA sequence such as promoters, non-coding sequences and randomly fragmented DNA. It is used in a wide array of biological experiments and practical applications ranging from genetic fingerprinting to large scale protein production.