Tuesday, October 18, 2016

You Can Clone Up To 5 Fragments In a Single Reaction


This week, we are highlighting the Gibson Assembly® HiFi 1-Step Kit, our quickest and easiest cloning solution. SGI-DNA offers the Gibson Assembly® HiFi 1-Step kit, Ultra kits, and Site-Directed Mutagenesis kits. The Gibson Assembly HiFi kit and Ultra kits are ideal for the assembly of plasmids and BAC constructs. While these kits both enable the cloning of multiple fragments, the numbers of fragments, the recommended sizes of the fragments, and the specifics of the workflow are different. Here, we’ll focus on the features of the HiFi kit.

The Gibson Assembly® HiFi 1-Step method allows for the isothermal assembly of up to 5 unique fragments. As implied, the HiFi 1-step process is performed in a single step. 


Gibson Assembly HiFi Protocol

To perform the HiFi 1-Step method, prepare DNA fragments and your vector of choice with overlapping ends. Combine 10-100 ng of each DNA fragment and the vector in equimolar amounts*, then add the HiFi mastermix. After incubating the reaction components at 50oC for 1 hour, the fully ligated constructs are ready to transform into competent cells. 

*If insert to be added is >1 kb, a 5-fold molar excess is suggested.




The Gibson Assembly® HiFi 1-Step kit is simple to use and ideal for assembling up to 5 fragments ranging in size from 500 bp to 32 kb. Try our 5 reaction HiFi starter kit today! It's the perfect introduction if you’re new to Gibson Assembly cloning. Larger kit sizes, including bulk pricing, are also available.

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