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Ligation-independent cloning (LIC) is a form of molecular cloning that can be performed without the use of restriction endonucleases or DNA ligase. The technique was developed in the early 1990s as an alternative to restriction enzyme/ligase cloning. [ 1 ]
Ligation is complicated by the fact that the reaction can involve both inter- and intra-molecular reactions, but the desired ligation products in many ligation reactions (e.g. ligating a DNA fragment into a vector) needed first to be inter-molecular, i.e. between two different DNA molecules, followed by an intra-molecular reaction to seal and ...
Polymerase cycling assembly (or PCA, also known as Assembly PCR) is a method for the assembly of large DNA oligonucleotides from shorter fragments. The process uses the same technology as PCR, but takes advantage of DNA hybridization and annealing as well as DNA polymerase to amplify a complete sequence of DNA in a precise order based on the single stranded oligonucleotides used in the process.
Molecular cloning generally uses DNA sequences from two different organisms: the species that is the source of the DNA to be cloned, and the species that will serve as the living host for replication of the recombinant DNA. Molecular cloning methods are central to many contemporary areas of modern biology and medicine. [2]
Golden Gate assembly involves digesting DNA sequences containing a type IIS restriction enzyme cut site and ligating them together. Golden Gate Cloning or Golden Gate assembly [1] is a molecular cloning method that allows a researcher to simultaneously and directionally assemble multiple DNA fragments into a single piece using Type IIS restriction enzymes and T4 DNA ligase. [2]
JBS USA and Perdue Farms will each pay $4 million for employing children through third-party staffing agencies, officials announced this week.
Other similar overlap-based assembly methods include Circular Polymerase Extension Cloning (CPEC), Sequence and Ligase Independent Cloning (SLIC) and Seamless Ligation Cloning Extract (SLiCE). [ 43 ] [ 44 ] [ 45 ] Despite the presence of many overlap assembly methods, the Gibson assembly method is still the most popular. [ 46 ]
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