Factors that affect Restriction Enzyme Activity The digestion activity of restriction enzymes depends on the following factors: Temperature: Most endonucleases digest the target DNA at 37°C with few exceptions. Methylation status of DNA: Methylation of adenine or cytidine residues affects the digestion of DNA.Click to see full answer. In respect to this, what is the mechanism of action of restriction enzymes?A major protective strategy for the host is to use restriction endonucleases (restriction enzymes) to degrade the viral DNA on its introduction into a cell. These enzymes recognize particular base sequences, called recognition sequences or recognition sites, in their target DNA and cleave that DNA at defined positions.Secondly, how do you know which restriction enzyme to use? When selecting restriction enzymes, you want to choose enzymes that: Flank your insert, but do not cut within your insert. Are in the desired location in your recipient plasmid (usually in the Multiple Cloning Site (MCS)), but do not cut elsewhere on the plasmid. Keeping this in view, what is the function of a restriction enzyme what determines the sites where a restriction enzyme works? A restriction enzyme is a protein that recognizes a specific, short nucleotide sequence and cuts the DNA only at that specific site, which is known as restriction site or target sequence. More than 400 restriction enzymes have been isolated from the bacteria that manufacture them.What are some common restriction enzymes?The most common Type II enzymes are those like HhaI (NEB #R0139), HindIII (NEB #R0104), and NotI (NEB #R0189), that cleave DNA within their recognition sequences.