Gateway® Recombination Cloning Technology
- 99% cloning efficiency delivers the clone you need
- Maintain orientation and reading frame throughout cloning process
- Efficient cloning of single fragments into multiple vectors simultaneously
- Flexibility to clone multiple gene fragments into a single construct
There are various methods to enter the Gateway® platform, including use of TOPO® cloning vectors containing Gateway® att sites, or purchasing an Ultimate™ ORF Clone already inserted into a Gateway® vector.
The typical cloning workflow, such as traditional restriction enzyme cloning, involves many steps, and can limit your cloning success. For example, certain restriction enzymes cannot be used because they might cut within your gene of interest, truncating the insert and making the gene useless for downstream expression. Additional clean-up steps are needed with this method, as you experience low-efficiency recovery of recombinants from cloning large fragments, and you waste time screening colonies to find the clone you need. All of these steps take considerable time and effort, and success is not guaranteed.
In contrast, Gateway® recombination cloning technology circumvents these cloning limitations, enabling you to access virtually any expression system.
Gateway® Technology - Meet the Inventor Series
Gateway® recombination cloning uses a one hour, 99%-efficient, reversible recombination reaction, without using restriction enzymes, ligase, subcloning steps, or screening of countless colonies, thereby saving you time, money, and effort. Widely adopted in the research community with more than 1,500 references since its launch, Gateway® technology makes collaboration across research disciplines easy and convenient and enables access to a multitude of vectors from these research groups for truly multidisciplinary scientific studies. Finally, new advancements such as MultiSite Gateway® Technology makes Invitrogen’s Gateway® cloning the ideal cloning method for protein expression and functional analysis.
Advantages of Gateway® Technology
- Fast, one hour, room temperature cloning reactions with >99% efficiency deliver the clone you need
- Maintaining orientation and reading frame without using restriction enzymes or ligation makes expression-ready clones
- Eliminating re-sequencing ensures consistent results throughout your experiment using the same clone from target identification to validation
- Shuttling insert DNA from one expression vector to another affords flexibility while simplifying your cloning workflow
Figure 1. Gateway® technology facilitates cloning of genes, into and back out of, multiple vectors via site-specific recombination. Once a gene is cloned into an Entry clone you can then move the DNA fragment into one or more destination vectors simultaneously.