Difference between revisions of "Team:Groningen/Results"
| Line 52: | Line 52: | ||
<li> <b>Construction</b><br> | <li> <b>Construction</b><br> | ||
<img class="imglabh" style="width: 35%; display:inline-block; float:right; margin-bottom:10px;" src="https://static.igem.org/mediawiki/2017/c/cc/24-10_DcAS9%2Bvrer_submission_gel_marked.png" alt="dCas9 colony PCR"> | <img class="imglabh" style="width: 35%; display:inline-block; float:right; margin-bottom:10px;" src="https://static.igem.org/mediawiki/2017/c/cc/24-10_DcAS9%2Bvrer_submission_gel_marked.png" alt="dCas9 colony PCR"> | ||
| − | For this part we started by making a biobrick compatible version of dCas9 called pSB1C3:dCas9QC. This part was also used for the construction of dCas9VRER. The full length of this part was sequenced (figure ..) and the results confirmed that the sequence was correct. In the notebook (week 18-09) a gel is shown from which it can be seen that the EcoRI site was successfully removed from our dCas9 part. | + | <p class="left">For this part we started by making a biobrick compatible version of dCas9 called pSB1C3:dCas9QC. This part was also used for the construction of dCas9VRER. The full length of this part was sequenced (figure ..) and the results confirmed that the sequence was correct. In the notebook (week 18-09) a gel is shown from which it can be seen that the EcoRI site was successfully removed from our dCas9 part.</p> |
<br> | <br> | ||
<b>Add sequencing image and find a way to incorporate dowloadablle sequence results</b> | <b>Add sequencing image and find a way to incorporate dowloadablle sequence results</b> | ||
<br> | <br> | ||
| − | Although the pSB1C3 | + | <p class="left">Although the pSB1C3:dCas9QC is already in the pSB1C3 backbone and contains all the correct restriction sites, it does not contained the suffix. To fix this mistake we restricted the pSB1C3dCas9 with EcoRI and SpeI and ligated it into pSB1C3, which was linearized with primers G69 and G70 (see notebook) and restricted with the same enzymes. In the gel on the right lane 2 & 3 correspond to the isolated plasmid restricted with respectively EcorI and a combination of EcorI and PstI. The sizes correspond to the linearized (6,2 kb) and the separate backbone (2 kb) and dCas9 part (4,2 kb).</p> |
| − | + | <p class="left">Since the chances of mutations in restriction-ligation cloning are negligible we did not fully sequence the part again. We did use pJet_Fwd and pJet-Rev to be absolutely certain that the correct part was cloned into the backbone. These results are shown in figure 3. </p> | |
Revision as of 15:09, 1 November 2017
Results
dCas9
- Construction
For this part we started by making a biobrick compatible version of dCas9 called pSB1C3:dCas9QC. This part was also used for the construction of dCas9VRER. The full length of this part was sequenced (figure ..) and the results confirmed that the sequence was correct. In the notebook (week 18-09) a gel is shown from which it can be seen that the EcoRI site was successfully removed from our dCas9 part.
Add sequencing image and find a way to incorporate dowloadablle sequence results
Although the pSB1C3:dCas9QC is already in the pSB1C3 backbone and contains all the correct restriction sites, it does not contained the suffix. To fix this mistake we restricted the pSB1C3dCas9 with EcoRI and SpeI and ligated it into pSB1C3, which was linearized with primers G69 and G70 (see notebook) and restricted with the same enzymes. In the gel on the right lane 2 & 3 correspond to the isolated plasmid restricted with respectively EcorI and a combination of EcorI and PstI. The sizes correspond to the linearized (6,2 kb) and the separate backbone (2 kb) and dCas9 part (4,2 kb).
Since the chances of mutations in restriction-ligation cloning are negligible we did not fully sequence the part again. We did use pJet_Fwd and pJet-Rev to be absolutely certain that the correct part was cloned into the backbone. These results are shown in figure 3.
- Experimental validation
After creating the part within the pSB1C3 biobrick format,.... - Considerations for replicating the experiments
Before this part can be fully integrated into a system, the next parameters should be measured:... - Future plans for the project
Based on these results we suggest that the next experiments can be conducted:...
dCas9 VRER
- Construction
For this part we started by.... - Experimental validation
After creating the part within the pSB1C3 biobrick format,.... - Considerations for replicating the experiments
Before this part can be fully integrated into a system, the next parameters should be measured:... - Future plans for the project
Based on these results we suggest that the next experiments can be conducted:...
CRISPR arrays
- Construction
For this part we started by.... - Considerations for replicating the experiments
Before this part can be fully integrated into a system, the next parameters should be measured:... - Future plans for the project
Based on these results we suggest that the next experiments can be conducted:...
Lactis toolbox
- Construction
For this part we started by.... - Experimental validation
After creating the part within the pSB1C3 biobrick format,.... - Considerations for replicating the experiments
Before this part can be fully integrated into a system, the next parameters should be measured:... - Future plans for the project
Based on these results we suggest that the next experiments can be conducted:...
hCas9 operon
- Construction
For this part we started by.... - Considerations for replicating the experiments
Before this part can be fully integrated into a system, the next parameters should be measured:... - Future plans for the project
Based on these results we suggest that the next experiments can be conducted:...
Reporter plasmid
- Construction
For this part we started by.... - Considerations for replicating the experiments
Before this part can be fully integrated into a system, the next parameters should be measured:... - Future plans for the project
Based on these results we suggest that the next experiments can be conducted:...
You should also describe what your results mean:
- Interpretation of the results obtained during your project. Don't just show a plot/figure/graph/other, tell us what you think the data means. This is an important part of your project that the judges will look for.
- Show data, but remember all measurement and characterization data must be on part pages in the Registry.
- Consider including an analysis summary section to discuss what your results mean. Judges like to read what you think your data means, beyond all the data you have acquired during your project.
Project Achievements
You can also include a list of bullet points (and links) of the successes and failures you have had over your summer. It is a quick reference page for the judges to see what you achieved during your summer.
- A list of linked bullet points of the successful results during your project
- A list of linked bullet points of the unsuccessful results during your project. This is about being scientifically honest. If you worked on an area for a long time with no success, tell us so we know where you put your effort.
Inspiration
See how other teams presented their results.
