ClonExpress II One Step Cloning (Vazyme, #C112) is a simple, fast, and high efficient cloning technology which is based on a homologous recombination technology. It 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 and incubated with Exnase II at 37°C for 30 min. The cloning products can then be directly transformed to competent cells with a true positive rate > 95%. This kit contains Exnase II and an optimized buffer with a unique recombinant enhancer which significantly improves the recombination efficiency. In addition, Exnase II is compatible with the reaction systems of restriction enzyme digestion and PCR, which means that both the digestion products of vector and the PCR products of insert can be directly used for recombination without purification, significantly simplifying the procedures.
Product Manual (pdf)

Advantages

• Easy, fast, and efficient.
• Directional cloning at any site on any vector.
• No need to consider the restriction enzyme cutting sites carried on inserts.
• Ligase-independent, true positive rate > 95%.
• Efficient cloning of fragments of 50 bp-10 kb.
• Linearized vector and PCR products can be used directly without purification.

When order, Cold Shipping option have to be selected, or your order may be delayed.

Selected Product Citations

1. Xing Y, et al. SLERT Regulates DDX21 Rings Associated with Pol I Transcription. Cell, 2017, 169(4):664-78. Pubmed ID: 28475895
2. Wu N, et al. TBX6 null variants and a common hypomorphic allele in congenital scoliosis. N Engl J Med, 2015, 4793:341-50. Pubmed ID: 25564734
3. Ge J, et al. Architecture of the mammalian mechanosensitive Piezo1 channel. . Pubmed ID: Nature, 2015, 527(7576):64-9. Pubmed ID: 26390154
4. Zhang XO, et al. Complementary Sequence-Mediated Exon Circularization. Cell, 2014, 159(1):134-47. Pubmed ID: 25242744
5. Zong Y, et al. Efficient C-to-T base editing in plants using a fusion of nCas9 and human APOBEC3A. Nature Biotechnology, 2018. Pubmed ID: 30272679
6. Li X, Wang Y, Liu Y, et al. Base editing with a Cpf1-cytidine deaminase fusion. Nature Biotechnology, 2018, 36(4):324-7. Pubmed ID: 29553573

Mechanism

Storage

All components should be Store at -20°C.

Components

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5. Mut Express II Fast Mutagenesis Kit V2 (Vazyme, #C214)
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