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Team:Macquarie Australia/Results
From 2013.igem.org
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Revision as of 11:29, 27 September 2013
Results and Characterisation
This page aims to provide a summary of our most important successful results, which have provided large strides towards the development of goal; Chlorophyll synthesis within E. coli. An in-depth view of our results can be found in our Notebook
BioBrick Construction
12 BioBricks were successfully constructed, a digest gel is shown below with bands corresponding to expected bp lengths for all genes.
Sequencing Results
Sequencing results obtained from our successful ligation of gBlock fragments
| BioBricks Constructed | Sequence Confirmed | Submitted to iGEM |
|---|---|---|
| POR |
 |
|
| ChlG |
|
|
| ChlP | | |
| ChlI2 | | |
| YCF54 | | |
| CTH1 | | |
| ChlI1 | | |
| Gun4 | | |
| Plastocyanin | | |
| ChlM | ||
| DVR1 | ||
| ChlD |
Characterisation
1st Attempt at ligation:
Promoter was ligated onto the genes and plated up on LB + CAM plates, we observed a lot of colony growth. We inoculated LB + CAM broth with a colony from the corresponding plate. A variety of growth was observed, a few plates corresponding to a certain genes had a lot more colonies than others, this can be observed from the image with the description order of genes. More colonies corresponding to a gene displayed more colony growth than others which can be observed from the image description showing the order of genes on the agar.
The PCR showed that the ligation was not successful, we derived the reason of this problem to either an improper clean up and therefore self ligation or improper restriction enzyme digest.
Attempt 2:
We re-attempted the ligation after a light change in the process, the vector was properly cleaned and restricted. The result was plated on LB + CAM plates, the results are to the left.
PCR of 2nd attempt:
The PCR showed that the ligation attempts were successful this time round, we exclaimed for joy. We used the BioBrick to inoculate E.coli .
