Difference between revisions of "Team:Groningen/Notebook"
| Line 41: | Line 41: | ||
<h5 id="snav1218">June 12-18</h5> | <h5 id="snav1218">June 12-18</h5> | ||
| − | <h6>General</h6> | + | <p> |
| − | + | <h6>General</h6> | |
| − | <h6>Interlab</h6> | + | This week we could finally start doing some real experiments in the lab! We started with optimizing the heat-shock time in the transformation protocol. We did this by transforming our competent cells with DNA from the competent cell test kit with various heat shock times (0-60 seconds). We found that the highest efficiency was obtained when using a heat shock of 30 seconds. |
| + | <h6>Interlab</h6> | ||
Because we did not have fully finished the design of any of the experiments for our own project yet, we started with the Interlab study. As a result we can proudly say that we were the first team to complete the Interlab project this year!! | Because we did not have fully finished the design of any of the experiments for our own project yet, we started with the Interlab study. As a result we can proudly say that we were the first team to complete the Interlab project this year!! | ||
We have successfully transformed all test devices of the interlab studies. Of each of the transformation plates one colony was picked, which was inoculated and grown overnight. After plasmid isolation of the ON cultures restriction analysis with EcoRI showed that all colonies contained the correct plasmids. In the remainder of the week we have completed the measurements according the plate reader protocol. More information about the interlab studies can be found on our <a href="https://2017.igem.org/Team:Groningen/InterLab">interlab page</a>. | We have successfully transformed all test devices of the interlab studies. Of each of the transformation plates one colony was picked, which was inoculated and grown overnight. After plasmid isolation of the ON cultures restriction analysis with EcoRI showed that all colonies contained the correct plasmids. In the remainder of the week we have completed the measurements according the plate reader protocol. More information about the interlab studies can be found on our <a href="https://2017.igem.org/Team:Groningen/InterLab">interlab page</a>. | ||
| Line 50: | Line 51: | ||
<h5 id="snav1925">June 19-25</h5> | <h5 id="snav1925">June 19-25</h5> | ||
<p> | <p> | ||
| + | <h6>General</h6> | ||
| + | We have inoculated one of the red colonies from the heatshock-test and isolated the plasmid (pSB1C3:bba_J04450) from it. This plasmid was used to confirm that the restriction enzymes, which were left over from the previous iGEM team of Groningen, worked properly. Moreover this plasmid was also used for cloning parts into the pSB1C3 backbone. | ||
| + | <h6>Lactis toolbox</h6> | ||
| + | After completing the interlab we started working on the lactis-toolbox. We started with two different parts of this sub-project. The first is the preparation of electro competent L. lactis NZ9000 cells and optimize the transformation protocols for it. The second was the preparation of a pTet-GFP/RFP and a pLac-GFP/RFP construct to test these promotors in L. lactis. From next week progress for this part of the project will be shown under the header <b>‘promotor testing’</b>. | ||
| + | This week we produced our first batch of competent lactis cells and determined a growth curve for NZ9000 in SGM17+glycine medium (see figure below). For the second part we successfully transformed E. coli with pLac (bba_), pTet (bba_), sfGFP (bba_) and a RBS (bba_). | ||
| + | <b>Insert figure lactis growth curve</b> | ||
| + | |||
</p> | </p> | ||
Revision as of 01:16, 1 November 2017
Notebook
Here you will find our complete labjournal. We documented our daily lab work per week.
June 05-11
Although we obtained the keys of our lab on the first of June we have not yet performed any real experiments. The entire week was spent on setting up the lab, e.g. getting all materials and machines needed. Also we made some media and standard buffers and we have produced our first batch of competent E. coli DH5 α at the end of this week.
June 12-18
General
This week we could finally start doing some real experiments in the lab! We started with optimizing the heat-shock time in the transformation protocol. We did this by transforming our competent cells with DNA from the competent cell test kit with various heat shock times (0-60 seconds). We found that the highest efficiency was obtained when using a heat shock of 30 seconds.Interlab
Because we did not have fully finished the design of any of the experiments for our own project yet, we started with the Interlab study. As a result we can proudly say that we were the first team to complete the Interlab project this year!! We have successfully transformed all test devices of the interlab studies. Of each of the transformation plates one colony was picked, which was inoculated and grown overnight. After plasmid isolation of the ON cultures restriction analysis with EcoRI showed that all colonies contained the correct plasmids. In the remainder of the week we have completed the measurements according the plate reader protocol. More information about the interlab studies can be found on our interlab page.June 19-25
General
We have inoculated one of the red colonies from the heatshock-test and isolated the plasmid (pSB1C3:bba_J04450) from it. This plasmid was used to confirm that the restriction enzymes, which were left over from the previous iGEM team of Groningen, worked properly. Moreover this plasmid was also used for cloning parts into the pSB1C3 backbone.Lactis toolbox
After completing the interlab we started working on the lactis-toolbox. We started with two different parts of this sub-project. The first is the preparation of electro competent L. lactis NZ9000 cells and optimize the transformation protocols for it. The second was the preparation of a pTet-GFP/RFP and a pLac-GFP/RFP construct to test these promotors in L. lactis. From next week progress for this part of the project will be shown under the header ‘promotor testing’. This week we produced our first batch of competent lactis cells and determined a growth curve for NZ9000 in SGM17+glycine medium (see figure below). For the second part we successfully transformed E. coli with pLac (bba_), pTet (bba_), sfGFP (bba_) and a RBS (bba_). Insert figure lactis growth curveJune 26-July 02
July 03-09
July 10-16
July 17-23
July 24-30
July 31-August 06
August 07-13
August 14-20
August 21-27
August 28-September 03
September 04-10
September 11-17
September 18-24
September 25-October 01
October 02-08
October 09-15
October 16-22
October 23-29
October 30-November 01
What should this page have?
- Chronological notes of what your team is doing.
- Brief descriptions of daily important events.
- Pictures of your progress.
- Mention who participated in what task.
Inspiration
You can see what others teams have done to organize their notes:
