Each of these methods have specific limitations ( Liang et al., 2017). To the best of our knowledge, of the many molecular cloning protocols that have been developed, the following are the main techniques currently used for routine cloning: restriction digestion- and ligation-based cloning ( Cohen et al., 1973), Gateway cloning ( Hartley et al., 2000), Gibson assembly ( Gibson et al., 2009), and Golden Gate cloning ( Engler et al., 2008). Thus, developing methods for the rapid and efficient construction of various vectors for transgenic research is more critical now than ever before. One of the main objectives in the post-genomics era is to functionally map gene expression data. Molecular cloning, which is one of the most fundamental procedures available for modern molecular biology research, has been critical for driving biotechnological advances. Due also to its simplicity and versatility, the cloning method has great potential for the modular assembly of DNA constructs. Nimble Cloning is applicable for the cloning of single or multiple fragments, as well as multi-site cloning. The cloning system is highly efficient, suitable for gene expression in both prokaryotic and eukaryotic expression systems, and enables the reuse of DNA fragments or plasmid entry clones.
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Both PCR products and plasmids can be used for the cloning reaction in the Nimble Cloning system.
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The cloning technique known as “Nimble Cloning” uses the restriction enzyme, SfiI, in combination with the T5 exonuclease, to linearize the vector and generate 3′-overhangs simultaneously. In this study, we have developed a novel method for standardized molecular cloning.
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Molecular cloning is one of the most fundamental technologies in molecular biology, and has been critical for driving biotechnological advances. Key Laboratory of Biology and Genetic Resources of Tropical Crops, Ministry of Agriculture, Institute of Tropical Bioscience and Biotechnology, Chinese Academy of Tropical Agricultural Sciences, Haikou, China.Pu Yan *, Yanjing Zeng, Wentao Shen, Decai Tuo, Xiaoying Li and Peng Zhou *