ExSembly™ Cloning Master Mix is developed and manufactured by Chesapeake Genomic Systems. ExSembly™ cloning technology enables rapid, single-step, high-efficiency homology-based insertion of any amplified DNA product into a CIRCULAR vector without prior DNA digestion step.
Chesapeake Genomic Systems
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| Mfr.No. | M0005 | ||||||||||||||||||||||||||||||||
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| Description | The ExSemblyTM cloning technology is simple, fast, and highly efficient, enabling directional insertion of any amplified DNA product into CIRCULAR vector. The key feature is that it completely eliminates the time-consuming step of preparing linear vector. The creative buffer system in our master mix allows restriction enzymes to efficiently digest the circular DNA and maintain high exonuclease and polymerase activity to enable the assembly to occur. This technology takes advantage of the fact that once the vector DNA successfully assembled with DNA inserts, the restriction sites in the vector disappears. It robustly removes background negative clones by completely digesting unassembled vector DNA. One can conveniently use a relatively large amount of vector DNA, up to 500 ng, and obtain a high number of positive clones, which consequently increases success rate.
Table 1. Comparison of assembling efficiency of ExSembly to assemble 5 fragments with those of other two competitors on the market, both of which require linearization and digestion of the DNA vector prior to the assembly. The ratio of all fragments was kept at 1:1. Both 500 ng and 1,000 ng of vector DNA were tried. The clones were plated on both ampicillin plates and ampicillin plus kanamycin plates. 32 colonies were picked for colony PCR screening, and the efficiencies were calculated and compared. |
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