Intact Genomics (IG®) propriety Quick10™ Cloning kit and DirectPlate® technologies enable simple, rapid and highly efficient cloning. The Quick10™ Cloning kit utilizes homologous assembly which allows for nearly 100% cloning accuracy. This kit enables the direct cloning of one or two DNA fragment(s) (<6kb total) through PCR into linearized cloning-ready Ig® AmpR/KanR-pCR vector (4 kb, provided in this kit). The PCR fragments can be generated by Intact Genomics high fidelity i7® DNA polymerase or other high fidelity DNA polymerases. Primers need to have 12 to 15 bases of homology at Ig® AmpR/KanR-pCR vector linear ends where the DNA fragment(s) or PCR product(s) needs to fuse. The unpurified PCR product (s) is then diluted 5-10 times by the dilution buffer included in this kit. No purification step is required.
The diluted PCR product (s) and linearized Ig® AmpR/KanR-pCR vector can then be assembled in just 7 minutes at 37 ºC, followed by a 3 minutes transformation step on ice utilizing our Directplate® XL DH10B competent cells (included in this kit). DirectPlate® cells eliminate heat shock, lengthy incubations, and time-consuming outgrowth procedures. Utilizing this kit, DNA/PCR product(s) can be assembled/transformed in 10 minutes vs. other seamless/fusion cloning, conventional T/A, or restriction ligation cloning and transformation methods that can take up to 2 hours or more.
