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Restriction digest is most commonly used as part of the process of the molecular cloning of DNA fragment into a vector (such as a cloning vector or an expression vector).The vector typically contains a multiple cloning site where many restriction site may be found, and a foreign piece of DNA may be inserted into the vector by first cutting the restriction sites in the vector as well the DNA ...
NdeI is a specific Type II restriction enzyme that cuts open specific target sequences, unlike exonucleases. [2] This enzyme is used in gene cloning to cut open reading frames in the plasmid of certain bacteria such as E. coli and insert a foreign gene, such as the gfpuv gene that codes for bio fluorescence of the jelly fish Aequorea victoria.
In molecular biology, XhoI is a type II restriction enzyme EC that recognise the double-stranded DNA sequence CTCGAG and cleaves after C-1. [1] Type II restriction endonucleases are components of prokaryotic DNA restriction-modification mechanisms that protect the organism against invading foreign DNA.
BglII catalyses phosphodiester bond cleavage at the DNA backbone through a phosphoryl transfer to water. [1] Studies on the mechanism of restriction enzymes have revealed several general features that seem to be true in almost all cases, although the actual mechanism for each enzyme is most likely some variation of this general mechanism.
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]
New England Biolabs (NEB) is an American life sciences company which produces and supplies recombinant and native enzyme reagents for life science research. [2] It also provides products and services supporting genome editing , synthetic biology and next-generation sequencing . [ 3 ]
NlaIII is a type II restriction enzyme isolated from Neisseria lactamica. [1] As part of the restriction modification system, NlaIII is able to prevent foreign DNA from integrating into the host genome by cutting double stranded DNA into fragments at specific sequences. [2]
Incomplete digestion by restriction enzymes after PCR can confound the analysis: incomplete digestion would suggest lack of DNA methylation (if cutting with a methylation-sensitive enzyme such as HpaII). It is also known that BstUI can cut at unconverted sites, leading to overestimation of methylation levels and so the use of HpaII is often needed.