The relationship between cloning efficiency and the length of overlapping ends.

<p>(<b>A</b>) Identification of positive and negative colonies by fluorescence. The circled colonies were non-fluorescent, which were used to calculate cloning accuracy of plasmid assembly by <i>E</i>. <i>coli in vivo</i> recombination. (<b>B</b>) The relationship between the number of colonies and overlapping nucleotides through 2-fragment assembly to construct pGFP. The data were from 2–6 independent experiments for each point, and the error bar represents the range of variation. (<b>C</b>) High cloning accuracy of pGFP 2-fragment assembly, regardless of the length of overlapping nucleotides. The 9 nt OL point had apparent 100% accuracy and no variation from 2 independent experiments, but it only had 7 colonies in total.