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The ability to synthesize DNA from an RNA template, via reverse transcription, enables researchers to study RNA with the same molecular approaches used for DNA investigations. cDNA generated by reverse transcription can be amplified using polymerase chain reaction (PCR). ...
A genomic library is a collection of the total genomic DNA from a single organism. The DNA is stored in a population of identical vectors, each containing a different insert of DNA. In order to construct a genomic library, the organism's DNA is extracted from cells and then digested with a restriction enzyme to cut the DNA into fragments of ...
Electroporation is a physical transfection method that uses an electrical pulse to create temporary pores in cell membranes through which substances like nucleic acids can pass into cells. It is a highly efficient strategy for the introduction of foreign nucleic acids into many cell types, including bacteria and mammalian cells. ...
In molecular biology, DNA ligase is a specific type of enzyme, a ligase, (EC 6.5.1.1) that facilitates the joining of DNA strands together by catalyzing the formation of a phosphodiester bond. It plays a role in repairing single-strand breaks in duplex DNA in living organisms, but some forms (such as DNA ligase IV) may specifically repair double-strand breaks (i.e. a ...
Agarose gel electrophoresis is a method of gel electrophoresis used in biochemistry, molecular biology, genetics, and clinical chemistry to separate a mixed population of DNA or proteins in a matrix of agarose. The proteins may be separated by charge and/or size (isoelectric focusing agarose electrophoresis is essentially size independent), and the DNA and RNA fragments by length ...
Agrobacterium tumefaciens, the cause of the economically important disease, crown gall, has also been studied for years because of its remarkable biology. The mechanism this bacterium uses to parasitize plant tissue involves the integration of some of its own DNA into the host genome resulting in unsightly tumors and changes in plant metabolism. A. tumefaciens prompted the first successful development ...
Many methods of delivering extra DNA into the nucleus of plant cells have been tried, and several have been successfully used to produce transgenic plants. The first to be discussed is the gene gun, also called particle acceleration or microprojectile bombardment. While this method was used to create several commercial events such as RoundupReady soybean, it has several limitations that ...
PCR (Polymerase Chain Reaction) is a revolutionary method developed by Kary Mullis in the 1980s. PCR is based on using the ability of DNA polymerase to synthesize new strand of DNA complementary to the offered template strand. Because DNA polymerase can add a nucleotide only onto a preexisting 3'-OH group, it needs a primer to which it can add the ...
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