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In 2012 a modified RAD tagging method called double digest RADseq (ddRADseq) was suggested. [10] [11] By adding a second restriction enzyme, replacing the random shearing, and a tight DNA size selection step it is possible to perform low-cost population genotyping.
The latter condition is of particular practical interest, since commercial restriction enzymes are usually supplied in a buffer containing a substantial amount of glycerol (50% v/v is typical), meaning insufficient dilution of the enzyme solution can cause star activity; this problem most often arises during double or multiple digests.
Different restriction enzymes acting on different recognition sites produce different DNA fragments. The term restriction enzyme originated from the studies of phage λ, a virus that infects bacteria, and the phenomenon of host-controlled restriction and modification of such bacterial phage or bacteriophage. [12]
Exonuclease III (ExoIII) is an enzyme that belongs to the exonuclease family. ExoIII catalyzes the stepwise removal of mononucleotides from 3´-hydroxyl termini of double-stranded DNA. [1]
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.
The enzyme degrades single-stranded DNA or RNA to nucleoside 5’-monophosphates, but does not digest double-stranded DNA, double-stranded RNA, or DNA / RNA hybrids. Mung Bean Nuclease catalyzes the specific degradation of single-stranded DNA or RNA, and produces mono and oligonucleotides carrying a 5′-P terminus.
HaeIII is one of many restriction enzymes (endonucleases) a type of prokaryotic DNA that protects organisms from unknown, foreign DNA. [1] It is a restriction enzyme used in molecular biology laboratories.
EcoRV (pronounced "eco R five") is a type II restriction endonuclease isolated from certain strains of Escherichia coli.It has the alternative name Eco32I. In molecular biology, it is a commonly used restriction enzyme.