The QuarCam Plus Seamless Cloning Kit is based on the Gibson Assembly principle and optimized for multi-fragment recombination, allowing seamless cloning of 1-12 insertion fragments (50 bp-12 kb) in a single run. The kit features optimized recombinase and buffer systems, and includes unique recombination enhancers to ensure high efficiency for both single and multiple fragment recombination. Additionally, it enhances the system's tolerance to impurities, enabling the direct use of linearized vectors and insertion fragments without purification, greatly simplifying the experimental process.

Seamless Cloning Principle

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Compatibility with Single and Multiple Fragment Homologous Recombination

Figure 1. Identification Results of Positive Clones with Insertion of Single or Multiple Fragments

Experiments were conducted using the QuarCam Plus Seamless Cloning Kit to recombine 1 and 4 fragments, respectively. The results demonstrated compatibility with the insertion of one or multiple target fragments.

- (A) Growth of E. coli cells with recombinant plasmids on antibiotic plates.

- (B) Agarose gel electrophoresis of colony PCR products for a single fragment insertion.

- (C) Agarose gel electrophoresis of colony PCR products for 4 fragment insertions.

- (D) Agarose gel electrophoresis of colony PCR products for 10 fragment insertions.

High Positive Clone Rate

Table 1. Statistics of Positive Clones with Insertion of Single or Multiple Fragments

Recombination experiments with insertion of 1, 4 or 10 fragments were conducted using products from Dynegene and a competitor, with 24 clones from each plate picked for colony PCR. Correct fragment size as shown by agarose gel electrophoresis was considered positive. Clones positive by colony PCR were further sequenced, with correctly sequenced recombination regions deemed positive. Results show higher positive rates for both single and multiple fragment recombination experiments with Dynegene products compared to the competitor.

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