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DNA ligase is an enzyme that joins together ends of DNA molecules. Although commonly represented as joining two pairs of ends at once, as in the ligation of restriction enzyme fragments, ligase can also join the ends on only one of the two strands (for example, when the other strand is already continuous or lacks a terminal phosphate necessary for ligation).
There are two common methods in which to construct a DNA molecular-weight size marker. [3] One such method employs the technique of partial ligation. [3] DNA ligation is the process by which linear DNA pieces are connected to each other via covalent bonds; more specifically, these bonds are phosphodiester bonds. [4]
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 ...
Amplified fragment length polymorphism (AFLP-PCR or AFLP) is a PCR-based tool used in genetics research, DNA fingerprinting, and in the practice of genetic engineering. Developed in the early 1990s by Pieter Vos, [1] AFLP uses restriction enzymes to digest genomic DNA, followed by ligation of adaptors to the sticky ends of the restriction ...
DNA ligase catalyzes the ligation of the 3' end of a DNA fragment to the 5' end of a directly adjacent DNA fragment. This mechanism can be used to interrogate a SNP by hybridizing two probes directly over the SNP polymorphic site, whereby ligation can occur if the probes are identical to the target DNA.
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]
The sheared DNA molecules are then subjected for the end repair and A-tailed treatment. The end repair treatment converts any damaged or incompatible protruding ends of DNA to 5’-phosphorylated and blunt-ended DNA, enabling immediate blunt-end ligation, while the A-tailing treatment adds an A to the 3’ end of the sheared DNA.
For example, DNA ligase can join two complementary fragments of nucleic acid by forming phosphodiester bonds, and repair single stranded breaks that arise in double stranded DNA during replication. In general, a ligase catalyzes the following dehydration reaction, thus joining molecules A and B: A-OH + B-H → A–B + H 2 O