Team:UofGuelph/Contributions
Registry Contributions
2018 Registry Submissions
Sequencing of BBa_K747047
Background
iGEM Guelph's 2018 contribution to the registry was the sequence analysis of a BioBrick located in the 2018 Distribution Kit.
BioBrick K747047 was located in Plate 2, Well 12O. Our team chose to sequence this part as the Distribution Kit listed it as a Bad Sequence and for our team’s first characterization of a Registry Part, we thought that this part was a good candidate. Sequencing preparation was performed by our team, while the sequencing was performed at the Laboratory Services at the University of Guelph.
Methods
A PCR was set up according to the following recipe:
| Component | Volume (µL) |
|---|---|
| H2O | 20.4 |
| Phusion Buffer | 8.0 |
| dNTPs | 0.8 |
| VF2 | 0.2 |
| VR | 0.2 |
| BBa_DNA | 10 |
| Phusion | 0.4 |
| Total | 40 |
PCR Cycling Parameters:
| Step | Temperature (°C) | Time (minutes) |
|---|---|---|
| Initial Denaturation | 98 | 2 |
| Denaturation | 98 | 1 |
| Annealing | 60 | 1 |
| Extension | 72 | 0.5 |
| Final Extension | 72 | 10 |
