Revision as of 03:19, 18 October 2018

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

"Denaturation", "Annealing", and "Extension" steps were repeated 34 times.

10µL of the PCR reaction was then run on a 1% agarose gel at 130V for 10 minutes to confirm presence of an amplicon.

The samples were then sent for sequencing via Sanger Sequencing.

10µL of the PCR reaction was then run on a 1% agarose gel at 130V for 10 minutes to confirm presence of an amplicon.

Results

Raw Sequence Data (trace files) can be found through this link

The sequences were then uploaded to the registry as part of our submission for our team to earn the Bronze Medal.