Description
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%.
Features
- Simple: Seamlessly assemble and clone up to six DNA fragments in a single reaction.
- Flexible: Design guidelines allow assembly into any vector of your choice.
- Efficient: Efficient for ligation of one to six fragments.
Application
- Quick Clone: It allows multiple overlapping DNA fragments to be seamlessly linked in a one-step, 15–60 minute isothermal reaction.
Specifications
| Cloning Process | Seamless cloning |
| Control | Positive control |
| Segments | Up to 6 fragments |
| Product Type | Seamless Cloning and Assembly Kit |
Components
| 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 |
Storage
The Hieff Clone™ Universal One Step Cloning Kit should be stored at -25℃~ -15°C for 1 year.
Figures
Figure 1 Multi-segment linkage: performance comparison with competitive strains (plate colony).
- Top half of the picture: 5 fragments+vector; Total segment length: 4000 bp; Vector length: 5000 bp
- Bottom half of the picture: 6 fragments+vector; Total segment length: 5000 bp; Vector length: 5000 bp
Figure 2 Multi-segment linkage: performance comparison with competitive strains (colony PCR).
- Top half of the picture: 5 fragments+vector; Total segment length: 4000 bp; Vector length: 5000 bp
- Bottom half of the picture: 6 fragments+vector; Total segment length: 5000 bp; Vector length: 5000 bp
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