Additional file 1: of Gibson Deletion: a novel application of isothermal in vitro recombination

The sequences of the oligonucleotides used in this study are reported. (XLSX 58 kb

Additional file 4: Figure S3. of Gibson Deletion: a novel application of isothermal in vitro recombination

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 clone’s 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 15481 kb

Additional file 5: Figure S4. of Gibson Deletion: a novel application of isothermal in vitro recombination

Sequences of the incorrect Gibson Deletion of 200 nt related to Fig. 5. Sequences of the DNA flanking the RFP cassette (red box) cloned in pUC19 using Gibson Deletion. The RFP cassette was PCR amplified using primers with homology arms aimed to delete 200 nts (100 nucleotides from each side of a KpnI cut) from the pUC19 plasmids (Fig. 5). All the clones picked and sequences had incorrect assembly and are presented here together with the predicted sequence on top (correct). Underlined sequences are part of the RFP cassette; red sequences are sequences that are deleted after assembly using Gibson Deletion; sequences in bold are insertions obtained after assembly. (PDF 1358 kb

Additional file 3: Figure S2. of Gibson Deletion: a novel application of isothermal in vitro recombination

Diagnostic cut related to Fig. 3. DNA isolated from clones that grew on selection plates was cut with the indicated enzymes. Group 1 cloning is depicted in Fig. 3a top panels and Group 2 cloning is depicted in Fig. 3a bottom panels. Images of simulated agarose gels are presented on the left of each cloning group. Red arrowheads indicate the clones with DNA that underwent successful/correct Gibson Deletion reaction. (PDF 3775 kb