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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]
This is the main advantage of OE-PCR and other long-homology based cloning methods such as Gibson assembly, which overcome the limitations of traditional restriction enzyme digestion and ligation cloning methods. [3] Assembly of custom DNA sequences with OE-PCR consists on three main steps.
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 ...
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]
Gibson assembly is a molecular cloning method that allows for the joining of multiple DNA fragments in a single, isothermal reaction. It is named after its creator, Daniel G. Gibson, who is the chief technology officer and co-founder of the synthetic biology company, Telesis Bio.
Restriction enzymes can generate a wide variety of ends in the DNA they digest, but in cloning experiments most commonly-used restriction enzymes generate a 4-base single-stranded overhang called the sticky or cohesive end (exceptions include NdeI which generates a 2-base overhang, and those that generate blunt ends). These sticky ends can ...
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.
HindIII (pronounced "Hin D Three") is a type II site-specific deoxyribonuclease restriction enzyme isolated from Haemophilus influenzae that cleaves the DNA palindromic sequence AAGCTT in the presence of the cofactor Mg 2+ via hydrolysis.