Fig. 2

Testing Gibson Deletion. a pUC19 plasmid was cut with KpnI or Acc65I and these restriction sites present at the 5′ end of the β-galactosidase gene were substituted with an AflII restriction site using Gibson Deletion. The red and yellow boxes represent the homologous sequences of 20 nucleotides each. A picture of blue (likely successful Gibson Deletion) and white (unsuccessful Gibson Deletion) bacteria colonies transformed with Gibson Deletion products is shown. b In the left schemes, the DNA added to each Gibson Deletion reactions are shown. The table reports the number of white or blue colonies after transformation and plating of each Gibson Deletion reaction. The number of correct clones after diagnostic cut and Sanger sequencing is reported in the last columns. The results of two independent experiments are shown