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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 ...
RRBS uniquely uses a specific restriction enzyme to enrich for CpGs. MspI digestion, or any restriction enzyme that recognizes CpG's and cuts them, produces only fragments with CG’s at the end. [2] This approach enriches for CpG regions of the genome, so it can decrease the amount of sequencing required as well as decrease the cost. [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]
The first few steps of COBRA, and the molecular changes caused by each step to methylated and unmethylated CpG sites. Combined Bisulfite Restriction Analysis (or COBRA) is a molecular biology technique that allows for the sensitive quantification of DNA methylation levels at a specific genomic locus on a DNA sequence in a small sample of genomic DNA. [1]
Digestion of total cellular DNA with one or more restriction enzymes and ligation of restriction half-site specific adaptors to all restriction fragments. Selective amplification of some of these fragments with two PCR primers that have corresponding adaptor and restriction site specific sequences.
Type I restriction enzymes possess three subunits called HsdR, HsdM, and HsdS; HsdR is required for restriction digestion; HsdM is necessary for adding methyl groups to host DNA (methyltransferase activity), and HsdS is important for specificity of the recognition (DNA-binding) site in addition to both restriction digestion (DNA cleavage) and ...
Thus, T-RFLP is different from ARDRA and RFLP in which all restriction fragments are visualized. In addition to these steps the TRFLP protocol often includes a cleanup of the PCR products prior to the restriction and in case a capillary electrophoresis is used a desalting stage is also performed prior to running the sample.
The density of RAD tags in a genome depends on the restriction enzyme used during the isolation process. [5] There are other restriction site marker techniques, like RFLP or amplified fragment length polymorphism (AFLP), which use fragment length polymorphism caused by different restriction sites, for the distinction of genetic polymorphism ...