Diagnostic cut related to Fig. 4. DNA isolated from blue clones was cut with KpnI or AflII and run on an agarose gel. The expected correct band pattern is reported in the inset on the right. The expected number of nucleotides deleted by Gibson Deletion (smaller deletions on top and larger deletions on the bottom) is reported on top of each group of lanes. Uncut, AflII or KpnI cut pUC19 are run on the left lanes of each gel as reported. Each cloneâs DNA was cut with AflII or KpnI and the two digestions run on consecutive lanes. DNA resulting from an incorrect assembly is labelled with a X on the bottom of the two lanes corresponding to the incorrect clone. (PDF 15481Â kb