Compatibility with Single and Multiple Fragment Homologous Recombination

 

Figure 1. Identification Results of Positive Clones with Insertion of 1 or 4 Fragments

 

Experiments were conducted using the QuarCam 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.

 

High Positive Clone Rate

 

Recombination System	Number of Inserted Fragments	Total Clones	Colony PCR Positive Rate	Sequencing Positive Rate	Overall Positive Rate
Dynegene	1	988	24/24 (100.0%)	19/20 (95.0%)	95.0%
	4	602	22/24 (91.7%)	16/20 (80.0%)	73.4%
Competitor	1	920	24/24 (100.0%)	15/20 (75.0%)	75.0%
	4	304	18/24 (75.0%)	11/20 (55.0%)	41.3%

 

Table 1. Statistics of Positive Clones with Insertion of 1 or 4 Fragments

 

Recombination experiments with insertion of 1 or 4 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.