b'PCR cloning PCR cloning is a method in which double-stranded DNA fragments amplified by PCR are ligated into a vector. With PCR amplification, this cloning technique requires much less starting material for the insert sequence and allows introduction of new restriction and/or recombination sites to the 5 end of the inserts.TOPO cloning Gateway cloningTOPO PCR cloning technology was developed to help improve cloningTo shuttle a PCR insert among vectors, the Gateway cloning system offers efficiency, simplify protocol setup, and accommodate a wide range ofsite-specific, recombinase-based cloning. It maintains the inserts proper PCR insert sizes. TOPO cloning vectors are linearized by the activity oforientation and reading frame during shuttling using the Gateway vectors. topoisomerase I (which also has a ligase function) that is covalently boundOnce a gene is cloned into an entry clone, you can then move the DNA to the 3 phosphate on each end (see figure below). This system enables thefragment into one or more destination vectors simultaneously.vectors to readily be joined to PCR inserts with compatible ends (with up to 95% efficiency), without the need for additional ligation steps, in 5 minutes. Find out more at thermofisher.com/gatewayFind out more at thermofisher.com/topoAdd 1 L of PCRPerform transformation Perform PCR with Taq orreaction to 1 L ofIncubate 5 minutes at with provided competent proofreading polymerase TOPO cloning vector room temperature E. coli strainA PCR product A PCR productCACC PCR productTOPOTOPOTOPOcloningvectorTOPO PCR cloning requires just three easy steps.CloningDid you know?The Invitrogen TOPO XL-2 Complete PCR Cloning Kit provides all the necessary elements for highly efficient cloning of extra-long PCR products from 113 kb. thermofisher.com/topoxl2 42'