b'OverviewPCR is a popular technique used in the preparation of synthetic genes. A double-stranded DNA fragment is amplified via PCR and ligated into a cloning vector. With PCR amplification, much less starting material is needed to make a cloning insert. New restriction and recombination sites can also be introduced at the 5 ends of inserts to facilitate cloning. Cloning genes longer than 2 kb or cloning more complex genes may require assembly of smaller DNA fragments, which can be obtained through PCR or construction of synthetic DNA fragments called Strings fragments.Obtain insertSelect vectorDesign PCRPerformVerify PCR for desiredprimers Perform PCR restrictionproduct Ligate Transform Select and purifyapplication enzyme digestionInvitrogen TOPONA Agarose gels NAcloning vectorInvitrogenHigh-fidelity PCRInvitrogenPlasmid OligoPerfect enzymes and E-GelPCR purificationCompetent cells preparation Primer Designer master mixes Thermo ScientificPower Snapkits and automationRestriction enzyme FastDigestElectrophoresis based cloning vector System Ligation master restriction enzymes mix and cloning enzymesInvitrogen GeneArt Strings DNA Fragments Gibson Assembly cloning or type IIs restriction enzyme digestion for larger or more complex genesInvitrogen GeneArt gene synthesis workflowCheck out our vector selection guide at thermofisher.com/strings 4 Contents'