b'Restriction enzyme cloning Found naturally in bacteria, restriction enzymes recognize and cleave specific DNA sequences, resulting in sticky ends (5 or 3 protruding ends) or blunt ends, enabling DNA inserts to be cloned into vectors with compatible ends. Star activity, buffer compatibility, and varying protocols for complete digestion are some common hurdles in restriction digestion.FastDigest restriction enzymes A BTo simplify cloning, we offer FastDigest enzymesan advanced line of restriction enzymes that share buffer compatibility with downstreamPlasmid DNA digested with FastDigest enzymes Plasmid DNA digested with NEB enzymes1 2 3 4 5 6 7 8 9 1 2 3 4 5 6 7 8 9modifying enzymes. Its benefits include: Complete digestion in 515 min Double and multiple digestions in a universal buffer for any combination of enzymes No sequential digestions and buffer changes176 unique specificities 1: Undigested plasmid DNA1: Undigested plasmid DNA 2: FastDigest BcuI2: SpeI-HF3: FastDigest XbaI3: XbaIDirect loading of reaction mixture on gels 4: FastDigest NdeI4: NdeI 5: FastDigest SalI5: SalI-HF 6: FastDigest XmaJI6: AvrII Find out more at thermofisher.com/fastdigest 7: FastDigest Eco31I7: BsaI 8: FastDigest Eco52I8: EaqI-HF 9: FastDigest BglII 9: BglIComparison of digestion efficiencies of restriction enzymes. (A) FastDigest restriction enzymes digest plasmid DNA much more efficiently compared to the (B) NEB enzymes. In this experiment, using the NEB protocol, 1 g of plasmid DNA was digested in ~15 min. Cloning Incomplete digestion is shown in red boxes.40'