Gene isolation and manipulation
35 important questions on Gene isolation and manipulation
The production of many DNA copies from one master region of DNA.
A protein (immunoglobulin) molecule, produced by the immune system, that recognizes a particular substance (antigen) and binds to it.
A pattern of dark spots in a developed photographic film or emulsion in the technique of autoradiography
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An F plasmid engineered to act as a cloning vector that can carry large inserts.
A library composed of cDNAs, not necessarily representing all mRNAs.
A method for the dissection of large segments of DNA, in which a cloned segment of DNA, usually eukaryotic, is used to screen recombinant DNA clones from the same genome bank for other clones containing neighboring sequences.
DNA transcribed from a messenger RNA template through the action of the enzyme reverse transcriptase.
The most popular method of DNA sequencing. It uses dideoxynucleotide triphosphates mixed with standard nucleotide triphosphates to produce a ladder of DNA strands whose synthesis is blocked at different lengths. This method has been incorporated into automated DNA-synthesis machines. Also called Sanger sequencing after its inventor, Frederick Sanger.
A section of DNA that has been inserted into a vector molecule, such as a plasmid or a phage, and then replicated to produce many copies.
An important enzyme in DNA replication and repair that seals the DNA backbone by catalyzing the formation of phosphodiester bonds.
A segment of DNA in which both strands have the same nucleotide sequence but in antiparallel orientation.
The collective techniques for obtaining, amplifying, and manipulating specific DNA fragments.
Any DNA to be used in cloning or in DNA-mediated transformation.
In a transgenic organism, the insertion of an introduced gene at a site other than its usual locus.
A vector that can carry a 35- to 45-kb insert of foreign DNA.
The use of a cloned fragment of wild-type DNA to transform a mutant into wild type; used in identifying a clone containing one specific gene.
A method of molecular separation in which DNA, RNA, or proteins are separated in a gel matrix according to molecular size, with the use of an electrical field to draw the molecules through the gel in a predetermined direction.
The inactivation of a gene by either a naturally occurring mutation or through the integration of a specially engineered introduced DNA fragment. In some systems, such inactivation is random, with the use of transgenic constructs that insert at many different locations in the genome. In other systems, it can be carried out in a directed fashion. See also targeted gene knockout.
The insertion of a genetically engineered transgene in place of a resident gene; often achieved by a double crossover
The process of producing modified DNA in a test tube and reintroducing that DNA into host organisms.
The cloning and molecular characterization of entire genomes.
(1) To form a hybrid by performing a cross. (2) To anneal complementary nucleic acid strands from different sources.
The transfer of electrophoretically separated RNA molecules from a gel onto an absorbent sheet, which is then immersed in a labeled probe that will bind to the RNA of interest.
An in vitro method for amplifying a specific DNA segment that uses two primers that hybridize to opposite ends of the segment in opposite polarity and, over successive cycles, prime exponential replication of that segment only
The identification of the DNA sequences encoding a gene of interest based on knowledge of its genetic or cytogenetic map location.
Describes a situation in which the phenotypic influence of a gene is altered by changes in the position of the gene within the genome.
Labeled nucleic acid segment that can be used to identify specific DNA molecules bearing the complementary sequence, usually through autoradiography or fluorescence.
A novel DNA sequence formed by the combination of two nonhomologous DNA molecules.
An endonuclease that will recognize specific target nucleotide sequences in DNA and break the DNA chain at those points; a variety of these enzymes are known, and they are extensively used in genetic engineering.
A DNA fragment resulting from cutting DNA with a restriction enzyme.
The transfer of electrophoretically separated fragments of DNA from a gel to an absorbent sheet such as paper; this sheet is then immersed in a solution containing a labeled probe that will bind to a fragment of interest.
A circular plasmid of Agrobacterium tumifaciens that enables the bacterium to infect plant cells and produce a tumor (crown gall tumor).
A gene that has been modified by externally applied recombinant DNA techniques and reintroduced into the genome by germ-line transformation.
An organism whose genome has been modified by externally applied new DNA.
Sanger sequencing makes use of ddNTP that blocks DNA synthesis after incorporation. How does it block DNA synthesis?
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