Results
Fig. 1 and 2 shows the results of colony PCR of E. coli transformed to confirm whether the target DNAs genes ① and ② synthesized have been cloned into the vector BBa_J04450.
Fig. 1 Genes ①
Fig. 2 Genes ②
For Fig. 1, bands were confirmed in lanes 1, 2, 3, and 4 nearby the
target molecular weight of 720 bp.
For Fig. 2, bands were confirmed in lanes 1, 2, 3, and 4 nearby the
target molecular weight of 1400 bp.
Furthermore, sequencing was performed to confirm that the cloning was performed correctly. And we get results that genes ① was 100% identical to the target sequence, but we failed to identify our target sequence in genes ②.
A vector containing the promoter of the target gene ① (BBa_I0500, BBa_14032, BBa_I13453) fig. 3, 4 and 5 in order shows the results of colony PCR of transformed E. coli to confirm that they were successfully cloned.
Fig. 3 BBa_I0500
Fig. 4 BBa_I13453
Fig. 5 BBa_I14032
Fig. 3 shows no band in all lanes 2, 3, 4 and 5.
In Fig. 4, bands were confirmed in lanes 3, 4, and 5 nearby the
target molecular weight of 810 bp.
In Fig. 5, a band near to the target molecular weight of 765 bp was
confirmed in lanes 2, 3, and 4.
From the results shown above, we know that the plasmid we designed
was constructed succesfully.