IN-FUSION—A efficient enzyme-free connection technology

For the traditional molecular cloning technology, the target gene is amplified by PCR. And then the T-A cloning vector is constructed and transferred to competence E.coli. Next, target gene fragment and vector are digested by the same restriction enzyme. After gel-purify, insert fragment and vector are ligated by ligaseWe will spend 2-3 weeks to complete this process.This technique has many drawbacks,as follows:1.The multiple cloning site is make up of many restriction enzyme cleavage sites,which may be not always suitable for the target gene;2.Adding the restriction enzyme cleavage site during the gene cloning by the PCR, which may bring non-specific amplification; 3.If the target gene is too long , the effect for connection with target vector is not very well.;4.It is difficult to accomplish the vector construction on a large scale.

     The IN-FUSION technology become actuality when the IN-FUSION enzyme was found. This enzyme is able to non-specific distinguish the linear DNA molecule’s 5’,3’ terminals,and digest 16 base from 5’ to 3’terminal.The linear DNA molecule’s 5’,3’ terminals will become the cohesive ends.If the target gene and linear target vector own the same cohesive ends,they cloud get together becoming a complete plasmid by the process of DNA renaturation.The IN-FUSION technology compared to the traditional molecular cloning technology,there are many advantages as follows: 1.The ligation reaction don’t need T4 DNA ligase and the restriction enzyme cleavage site; 2.There will only spent 2-3 days finishing the process of molecular clone; 3.The connection efficiency is higher than the traditional molecular cloning technology about 200-500 times; 4.The long of gene fragment inseted the target vector will become much easier;5.It is suit for accomplishing the vector construction on a large scale.

Pic:IN-FUSION technical processes

The CLOUD-CLONE CROP have an efficient IN- FUSION enzyme system and support technology services.This sysytem is suit for kinds of gene. we also provide vector construction services.Welcome to consulting!

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