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Minipreparation of plasmid DNA is a rapid, small-scale isolation of plasmid DNA from bacteria. [20] [21] Commonly used miniprep methods include alkaline lysis and spin-column based kits. [3] [22] It is based on the alkaline lysis method. The extracted plasmid DNA resulting from performing a miniprep is itself often called a "miniprep".
Alkaline lysis is the process of isolating plasmid deoxyribonucleic acid (DNA) in bacteria. It is a standard method used in molecular biology to isolate the plasmid without obtaining chromosomal DNA. The first alkaline lysis was performed by Birnom and Doly in 1979. [ 1 ]
Structure of a lipopolysaccharide (LPS) Lipopolysaccharide, now more commonly known as endotoxin, [1] is a collective term for components of the outermost membrane of the cell envelope of gram-negative bacteria, such as E. coli and Salmonella [2] with a common structural architecture.
Atlantic horseshoe crab Limulus polyphemus. Limulus amebocyte lysate (LAL) is an aqueous extract of motile blood cells from the Atlantic horseshoe crab Limulus polyphemus.LAL reacts with bacterial endotoxins such as lipopolysaccharides (LPS), which are components of the bacterial capsule, the outermost membrane of cell envelope of gram-negative bacteria.
In this method, a fragment of DNA is synthesized, and then inserted into a plasmid. [14] It involves the cleavage by a restriction enzyme at a site in the plasmid and subsequent ligation of a pair of complementary oligonucleotides containing the mutation in the gene of interest to the plasmid. Usually, the restriction enzymes that cut at the ...
Plasmid Cloning Guides 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 ...