Hieff Clone™ Universal One Step Cloning Kit -10922ES

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YeasenSKU: 10922ES20

Size: 20T
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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%.


  • 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.


  • Quick Clone: It allows multiple overlapping DNA fragments to be seamlessly linked in a one-step, 15–60 minute isothermal reaction.


Cloning Process Seamless cloning
Control Positive control
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).

  • 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
Citations & References:

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[9] Yu Z, Shi M, Stewart T, et al. Reduced oligodendrocyte exosome secretion in multiple system atrophy involves SNARE dysfunction. Brain. 2020;143(6):1780-1797. doi:10.1093/brain/awaa110(IF:11.337)

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[12] Li N, Xu Y, Chen H, et al. NEIL3 contributes to the Fanconi anemia/BRCA pathway by promoting the downstream double-strand break repair step. Cell Rep. 2022;41(6):111600. doi:10.1016/j.celrep.2022.111600(IF:9.995)

[13] Tao R, Shi M, Zou Y, et al. Multicoloured fluorescent indicators for live-cell and in vivo imaging of inorganic mercury dynamics. Free Radic Biol Med. 2018;121:26-37. doi:10.1016/j.freeradbiomed.2018.04.562(IF:6.020)

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