Team:Copenhagen/Notebook
From 2012.igem.org
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| | The sequencing results have come back, and everything was as we hoped. | | The sequencing results have come back, and everything was as we hoped. |
| | It took more time than expected to get the new primers so the first days has been spend applying for sponsors, remaking our website, and deciding how to do the next experiments. | | It took more time than expected to get the new primers so the first days has been spend applying for sponsors, remaking our website, and deciding how to do the next experiments. |
| - | PCR on the backbone (pSB1C3), and our constitutive promoter has been unsuccessful while we have been able to amplify and cut out a fine band of YF1. | + | PCR on the backbone (pSB1C3), and our constitutive promoter (ProC) has been unsuccessful while we have been able to amplify and cut out a fine band of the gene encoding the histidine kinase YF1. |
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| - | We're still having trouble with both pSB1C3 and ProC. We're currently running a self-annealing programme for ProC because it is to small to do PCR. pSB1C3, on the other hand, seems too big so therefore we've lowered the annealing temperature and chosen a longer extension time. So far it has had no effect but we will keep optimizing. | + | This week, we managed to amplify most of the genes needed in our control construct! |
| | + | We succesfully amplified the promoter FixK2 and the gene encoding its regulator FixJ. |
| | + | We found out that the backbone, pSB1C3, was supplied as linear DNA and we discovered its conversion into circular DNA after transformation into E. coli. We confirmed the insertion of the backbone DNA by another discovery - red fluorescence when looking at the bacteria with fluorescence microscopy because the pSB1C3 biobrick contains a red flourescence protein gene - wow! We purified DNA from these bacteria and finally got our DNA product. |
| | + | We also managed to amplify the terminator sequence. |
| | + | We are having problems with PCR on the large luxCDABE cassette but we found out that the template we are using might be contaminated. We are also still having trouble with the very small constitutive promoter proC - we forund out that it is too small for PCR because it is about the same size as the primers.. So we have tried to amplify it using the gene in two single strands - which we have tried to anneal to each other - but this has been unsuccesful so far. |
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Revision as of 13:26, 13 July 2012
Notebook
Follow our work here in the notebook.
We will post the most important of our findings from the lab-work. We will also post pictures from our great summer!!! The light at the department of Plant Biology will probably shine brighter than the Danish summer sun anyway ;)
June
Week 1: 4th-8th
Exam period starts :(
Week 2: 11th-15th
Exams for a lot of our team members but we start designing our primers.
Week 3: 18th-22nd
Primers are designed and ordered so we'll have them next week.
Week 4: 25th-29th
Exams are finally over and we can start some of the lab work! Yay!!! :D
We've divided our construct into two halfs and will in this week be doing PCR on the first half of our construct. We've designed primers for the second half of our construct and ordered them.
All our templates, kindly provided by Uppsala, are sent to sequencing in Holland.
July
Week 1: 2nd-6th
The sequencing results have come back, and everything was as we hoped.
It took more time than expected to get the new primers so the first days has been spend applying for sponsors, remaking our website, and deciding how to do the next experiments.
PCR on the backbone (pSB1C3), and our constitutive promoter (ProC) has been unsuccessful while we have been able to amplify and cut out a fine band of the gene encoding the histidine kinase YF1.
Week 2: 9th-13th
 Info |
This week, we managed to amplify most of the genes needed in our control construct!
We succesfully amplified the promoter FixK2 and the gene encoding its regulator FixJ.
We found out that the backbone, pSB1C3, was supplied as linear DNA and we discovered its conversion into circular DNA after transformation into E. coli. We confirmed the insertion of the backbone DNA by another discovery - red fluorescence when looking at the bacteria with fluorescence microscopy because the pSB1C3 biobrick contains a red flourescence protein gene - wow! We purified DNA from these bacteria and finally got our DNA product.
We also managed to amplify the terminator sequence.
We are having problems with PCR on the large luxCDABE cassette but we found out that the template we are using might be contaminated. We are also still having trouble with the very small constitutive promoter proC - we forund out that it is too small for PCR because it is about the same size as the primers.. So we have tried to amplify it using the gene in two single strands - which we have tried to anneal to each other - but this has been unsuccesful so far.
Week 3: dato
JahDah JahDah ... Text ...
Week 4: dato
JahDah JahDah ... Text ...
August
Week 1: dato
JahDah JahDah ... Text ...
Week 2: dato
JahDah JahDah ... Text ...
Week 3: dato
JahDah JahDah ... Text ...
Week 4: dato
JahDah JahDah ... Text ...
September
Week 1: dato
JahDah JahDah ... Text ...
Week 2: dato
JahDah JahDah ... Text ...
Week 3: dato
JahDah JahDah ... Text ...
Week 4: dato
JahDah JahDah ... Text ...
October
Week 1: dato
JahDah JahDah ... Text ...
Week 2: dato
JahDah JahDah ... Text ...
Week 3: dato
JahDah JahDah ... Text ...
Week 4: dato
JahDah JahDah ... Text ...
This is our notebook, displaying how our work has progressed throughout the summer.
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