Hieff Clone™ Universal One Step Cloning Kit is a simple, fast, and highly efficient cloning technology and enables directional insertion of any amplified DNA product into any linearized vector at any site. Firstly, the vector is linearized at the cloning site. A small sequence overlapped with each end of the cloning site is added onto the insert through PCR. The insert and the linearized vector, with overlapped sequences of 15 bp - 20 bp on both 5' - and 3' -end, respectively, are mixed in an appropriate ratio and incubated with recombinase at 50℃ for 5-15 min.
Hieff Clone™ Universal one step cloning kit is a novel cloning Kit, independent of DNA ligase, significantly reducing the self-ligated colonies and bringing a true positive rate >95%.
|Cloning Process||Seamless cloning|
|Segments||Up to 6 fragments|
|Product Type||Seamless Cloning and Assembly Kit|
|Components No.||Name||10922ES20 (20T)|
|10922-A||Hieff Clone™ Universal Enzyme Premix||200 μL|
|10922-B||500 bp Control Insert (25 ng/μL)||5 μL|
|10922-C||pUC 19 Control Vector, linearized (50 ng/μL)||5 μL|
The Hieff Clone™ Universal One Step Cloning Kit should be stored at -25℃~ -15°C for 1 year.
Figure 1 Multi-segment linkage: performance comparison with competitive strains (plate colony).
Figure 2 Multi-segment linkage: performance comparison with competitive strains (colony PCR).
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