Invitrogen manual gateway

Gateway Cloning The easy-to-use choice for everyday cloning Invitrogen Gateway recombination cloning technology circumvents traditional restriction enzyme based cloning limitations, enabling you to access virtually any expression system in just a few simple steps.  · The GATEWAY BP Cloning Reaction The BP reaction is used to create an Entry Clone from Expression Clones (Figure 4) or PCR products (Figure 6). Once a gene is flanked by attL sites (Entry Clone), it can be transferred into any number of Destination Vectors to generate new Expression Clones. The Gateway cloning method, developed by Invitrogen, is an in vitro version of the integration and excision recombination reactions that take place when lambda phage infects bacteria. I n vivo, t hese recombination reactions are facilitated by the recombination of attachment sites from the phage (att P) and the bacteria (att B).

Invitrogen™ Gateway™ LR Clonase™ II enzyme mix is a proprietary enzyme and buffer formulation containing the bacteriophage lambda recombination proteins Integrase (Int) and Excisionase (Xis), the E. coli-encoded protein Integration Host Factor (IHF). Gateway® LR Clonase® II enzyme mix catalyzes the in vitro recombination between an entry clone (containing a gene of interest flanked by attL sites) and a destination vector (containing attR sites) to generate an expression www.doorway.ruy® LR Clonase® II contains enzymes and buffer in a single mix to enable convenient ten-microliter reaction set-up with fewer pipetting steps. Article Snippet: Gateway cloning To generate pENTR plasmids, sequences of interest flanked by the appropriate Gateway attB sites, as listed in the MultiSite Gateway Pro manual, were either synthesized as gBlocks or PCR amplified by Platinum Taq HiFi DNA polymerase (ThermoFisher) or Q5 High-Fidelity DNA Polymerase (NEB) and subsequently purified.

Invitrogen™ Gateway™ LR Clonase™ II enzyme mix is a proprietary enzyme and buffer formulation containing the bacteriophage lambda recombination proteins Integrase (Int) and Excisionase (Xis), the E. coli-encoded protein Integration Host Factor (IHF), and reaction buffer provided in a single mix for convenient reaction set up. Gateway™. Invitrogen Gateway recombination cloning technology circumvents traditional restriction enzyme based cloning limitations, enabling you to access virtually any expression system in just a few simple steps. Here is an archived copy of the Invitrogen 3-insert multisite Gateway manual, version D, We originally had a live link to the Invitrogen site, but after this link broke it seemed easier to hard-archive a copy.