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Cloning

MultiSite Gateway® Technology

What if you could easily and accurately assemble 2, 3 or even 4 DNA fragments in the order and orientation of your desire into one construct? Imagine reconstituting key element(s) of a pathway or functional complex within one plasmid. Now you can do all of this and more.
Compared to subcloning using restriction enzymes, Gateway recombination cloning technology is efficient, simple and effective, saving you time and effort (Table 1). Now, with added flexibility to assemble multiple DNA fragments precisely, efficiently, and directionally in a defined order and orientation and without subcloning, Multisite Gateway is the ideal choice for protein expression.

Table1 : Gateway® Recombination Cloning Simplifies Cloning Workflow

Steps Gateway® Recombination Cloning Sub-cloning with Restriction enzymes
Existing primers? Yes No
Vector ready for cloning? Yes No
Ligation reagents included? Yes No
Competent cells separately? Included Purchase separately: 0 hours
Prepare:  up to 6 hours
Vector clean up? No Yes
PCR fragment cleanup? No Yes
Recombination efficiency Up to 99% ~50%
Cloning time into expression vector* 65 minutes up to 24 hours

*excludes miniprep time common to both methods

Applications for MultiSite Gateway® Technology

Discover how MultiSite Gateway® Technology gives you opportunity to perform pathway reconstitution, multiple gene expression and regulation, protein interaction studies, and more. Easily swap and assemble promoters, tags, and genes for your applications, or build libraries of different elements for screening purposes (Table 2).


See Data Generated with MultiSite Gateway® Pro Kits


Table 2

Add promoter/tag elements to you Gateway® Entry clones
Combinatorial tagging
Construct biological or biochemical pathways
Expression of enzymatic pathways
Expression of multi-subunit protein complexes
Gene knock-down and rescue
Optimized multi-gene delivery without co-transfection
Screen expression from different promoters
Shuffle protein coding domains
Variable gene expression levels using different expression elements
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