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The blue–white screen is a screening technique that allows for the rapid and convenient detection of recombinant bacteria in vector-based molecular cloning experiments. This method of screening is usually performed using a suitable bacterial strain , but other organisms such as yeast may also be used.
This allows for blue–white screening when using host strains such as E. coli JM109, which produces only the C-terminal portion of lacZ, also known as the β-polypeptide. [3] If pUC19 is inserted into E. coli JM109 and grown on agar media supplemented with IPTG and X-gal , then colonies will appear blue, as the plasmid encodes for the α ...
DH5-Alpha Cells are E. coli cells engineered by American biologist Douglas Hanahan to maximize transformation efficiency. They are defined by three [1] mutations: recA1, endA1 which help plasmid insertion and lacZΔM15 which enables blue white screening.
The most widely used is the gene coding for E. coli β-galactosidase, whose activity can easily be detected by the ability of the enzyme it encodes to hydrolyze the soluble, colourless substrate X-gal (5-bromo-4-chloro-3-indolyl-beta-d-galactoside) into an insoluble, blue product (5,5'-dibromo-4,4'-dichloro indigo).
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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 ...
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