b'Restriction enzyme cloningRestriction enzymes that recognize and cleave specific DNA sequences are made naturally by bacteria. Cleavage can create 5 or 3 sticky ends or blunt ends that enableA BDNA inserts to be cloned into vectors with compatible ends. Star activity (nonspecificFastDigest enzymes Another vendors enzymescleavage), buffer incompatibility, and variation between protocols for complete digestion1 2 3 4 5 6 7 8 9 1 2 3 4 5 6 7 8 9are some common problems in the use of restriction digestion.PCR is commonly used to add restriction sites to inserts to facilitate cloning. Prior to selecting your restriction sites, it is important to obtain a restriction map of your gene to make sure the sites are not present in your insert. FastDigest restriction enzymes 1: Undigested plasmid DNA1: Undigested plasmid DNA We offer FastDigest restriction enzymes to help simplify cloning. FastDigest restriction2: FastDigest BcuI2: SpeI-HF3: FastDigest XbaI3: XbaI enzymes enable complete digestion in 15 minutes and work with a single buffer that4: FastDigest NdeI4: NdeI 5: FastDigest SalI5: SalI-HF is compatible with enzymes for downstream modification. Use our FastDigest Enzyme6: FastDigest XmaJI6: AvrII Selection Tool to find the right ones for your cloning needs, and view a protocol for rapid7: FastDigest Eco31I7: BsaI 8: FastDigest Eco52I8: EaqI-HF DNA digestion with FastDigest enzymes. 9: FastDigest BglII 9: BglITroubleshooting tip: restriction digestion reactions Comparison of restriction enzyme digestion efficiency. (A) FastDigest restriction enzymes digest plasmid DNA much more efficiently than enzymes from another vendor. In this experiment, 1 g of plasmid DNA was digested for approximately 15 minutes. (B) Incomplete digestion indicated by the red boxes.Find out more at thermofisher.com/fastdigest 6 Contents'