Here we show our PCR results, and the conditions used, for the fragments which we subsequently used in Gibson assembly, as well as the sequencing results for our part, NrfB.

PCR results

The following table shows the final conditions used for successful PCR:

An agarose gel of final fragment PCR products is shown below:

Sequencing results

We successfully performed PCR on our fragments with our editing primers, to isolate the whole coding regions. Sequencing of our constructs showed that most had significant deletions and insertions of unknown DNA, however one construct, NrfB pSB1C3, had only one point mutation (shown below). We submitted this part (see our Parts page), annotating the mutation so future teams can take it into consideration when ordering the part. The part can still be used accurately if this mutation is edited, either molecularly, or through DNA synthesis.