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TOOLS UltraFast PCR cloning kit is designed for fast and directional cloning of the single or multiple DNA into any vector at any site, regardless of DNA fragment end compatibility. This kit is applicable for efficient clone of fragments of 50 bp - 10 kb. The vector is linearized at the cloning site. A small sequence overlapped with each end of the cloning site is added onto the insert through PCR. The insert and the linearized vector, with overlapped sequences of 15 bp - 20 bp on both 5’- and 3’-end, respectively, are mixed in an appropriate ratio and incubated with recombinase 2X UltraFast Mix at 50°C for 5 - 15 min. This kit is a novel cloning kit, applicable for homologous recombination with 1 - 5 fragments. The Kit is independent of DNA ligase, significantly reducing the self-ligated colonies. The enhanced recombinase and highly optimized buffer included in 2X UltraFast Mix significantly improve the recombination efficiency and the tolerance to impurities. pFAST vector is compatible with most PCR products, enabling the specific PCR products to be used directly for recombination without any treatments, which significantly simplify the procedure.