Note that the Tol2kit is based on the original threepart multisite Gateway system (as described in the version D manual), in which destination vectors use attR4R3 sites, not the new Multisite Gateway Pro system, in which all the destination vectors use attR1R2 sites.
While the donor, entry, and destination vectors are incompatible with the MultiSite Gateway Pro Technology enables you to efficiently and conveniently assemble multiple DNA fragments in the desired order and orientation into a Gateway Expression vector.
Using specifically designed att sites for recombinational cloning, you can clone two, three, or four DNA fragments into Schematic diagram of threefragment Gateway MultiSite recombination cloning.
Fragments 1, 2, and 3 are amplified by PCR using protocols provided in the manual. BP Recombination Reaction Perform a BP recombination reaction between each attBflanked DNA fragment and the appropriate attPcontaining donor vector to generate an entry clone (see page 26 MultiSite Gateway Pro Plus Kit 20 reactions User Manual Corporate Headquarters Invitrogen Corporation 1600 Faraday Avenue Carlsbad, CA T: 1 760 603 7200 F: 1 760 602 6500 JumpIn TI Gateway Targeted Integration System MultiSite Gateway An advance in recombination cloning technology was the development of multifragment recombination cloning that allows simultaneous cloning of more than one DNA fragment into an expression vector (marketed as Gateway MultiSite recombination cloning by Invitrogen).
This multifragment recombination cloning system utilizes mutant Schematic modified from the Invitrogen MultiSite Gateway Pro user manual. Mentions: A schematic diagram of fourfragment Gateway MultiSite recombination cloning is shown in a MultiSite Gateway reaction (MultiSite GatewayPro, Invitrogen), genes or Gateway reaction according to the manufacturer's manual.
TOPO TA Cloning To Create a Gateway MultiSite Gateway Pro technology is the answer, allowing you the flexibility to assemble multiple DNA fragments precisely, efficiently, and directionally in a defined order and orientation and without sub cloning.
viii Kit Contents and Storage Types of Kits This manual is supplied with the kits listed below. Product Catalog no. MultiSite Gateway Pro 2. 0 Kit for 2fragment recombination MultiSite Gateway Pro 3. 0 Kit for 3fragment recombination MultiSite Gateway Pro 4. 0 Kit for 4fragment recombination MultiSite Gateway Pro This is a versatile cloning toolkit for constructing fullycustomizable threepart fusion proteins, based on the MultiSite Gateway cloning Multisite gateway pro manual from Life Technologies.
The fusion protein components (i. e. an ORF of interest plus two flanking modules) are encoded in the three plasmids integral to the original MultiSite Gateway kit.
MultiSite Gateway Pro Cloning. MultiSite Gateway Pro technology enhances the classic Gateway cloning protocol by allowing you to clone up to four regions at one time. In MultiSite Gateway Pro cloning, three additional types of recombination sites have been engineered to recombine uniquely between the two ends of a source sequence and a