Difference between revisions of "Team:Tartu TUIT/Notebook"
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<div class="event" data-event-date="06.04.2018" data-event-type=""> | <div class="event" data-event-date="06.04.2018" data-event-type=""> | ||
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| − | <p>Searched for the pathway of synthesis of mycosporine-like amino acids from the genes of different organisms. Actinosynnema mirum DSM with the gene names amir_4256, amir_4257, amir_4258, amir_4259. And Nostoc commune with the gene names mysA, mysB mysC and mysD.<img src="notebook1.jpg"></p> | + | <p>Searched for the pathway of synthesis of mycosporine-like amino acids from the genes of different organisms. Actinosynnema mirum DSM with the gene names amir_4256, amir_4257, amir_4258, amir_4259. And Nostoc commune with the gene names mysA, mysB mysC and mysD.<img src="https://static.igem.org/mediawiki/2018/8/80/T--Tartu_TUIT--notebook1.jpg"></p> |
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<div class="event" data-event-date="22.05.2018" data-event-type=""> | <div class="event" data-event-date="22.05.2018" data-event-type=""> | ||
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| − | <p>iGEM distribution kit arrived.<img src="notebook2.jpg"></p> | + | <p>iGEM distribution kit arrived.<img src="https://static.igem.org/mediawiki/2018/4/41/T--Tartu_TUIT--notebook2.jpg"></p> |
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<div class="event" data-event-date="20.06.2018" data-event-type=""> | <div class="event" data-event-date="20.06.2018" data-event-type=""> | ||
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| − | <p>Primers arrived.<img src="notebook3.jpg"></p> | + | <p>Primers arrived.<img src="https://static.igem.org/mediawiki/2018/d/dd/T--Tartu_TUIT--notebook3.jpg"></p> |
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<div class="event" data-event-date="21.06.2018" data-event-type=""> | <div class="event" data-event-date="21.06.2018" data-event-type=""> | ||
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| − | <p>Started the first cloning process Biobricks with PCR, followed by restriction and overnight ligation.<img src="notebook4.jpg"></p> | + | <p>Started the first cloning process Biobricks with PCR, followed by restriction and overnight ligation.<img src="https://static.igem.org/mediawiki/2018/5/51/T--Tartu_TUIT--notebook4.jpg"></p> |
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<div class="event" data-event-date="04.07.2018" data-event-type=""> | <div class="event" data-event-date="04.07.2018" data-event-type=""> | ||
<div class="details"> | <div class="details"> | ||
| − | <p>Team photoshoot. <img src="notebook5.jpg"></p> | + | <p>Team photoshoot. <img src="https://static.igem.org/mediawiki/2018/c/cc/T--Tartu_TUIT--notebook5.jpg"></p> |
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<div class="event" data-event-date="13.07.2018" data-event-type=""> | <div class="event" data-event-date="13.07.2018" data-event-type=""> | ||
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| − | <p>CPEC, gybson, vegas cloning initiation. PCR with Phusion Polymerase.<img src="notebook6. | + | <p>CPEC, gybson, vegas cloning initiation. PCR with Phusion Polymerase.<img src="https://static.igem.org/mediawiki/2018/2/23/T--Tartu_TUIT--notebook6.jpg"></p> |
<table> | <table> | ||
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<div class="event" data-event-date="19.07.2018" data-event-type=""> | <div class="event" data-event-date="19.07.2018" data-event-type=""> | ||
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| − | <p>-Skype conference with Ecuador team<img src="notebook7.jpg"></p> | + | <p>-Skype conference with Ecuador team<img src="https://static.igem.org/mediawiki/2018/5/52/T--Tartu_TUIT--notebook7.jpg"></p> |
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| − | <div class="actions"><a class="button" href="">Read the whole notebook in | + | <div class="actions"><a class="button" href="https://benchling.com/s/etr-gWRVXVpu9z2pgLl1GxAc">Read the whole notebook in our benchling</a></div> |
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</section> | </section> | ||
Latest revision as of 03:46, 18 October 2018
Searched for the pathway of synthesis of mycosporine-like amino acids from the genes of different organisms. Actinosynnema mirum DSM with the gene names amir_4256, amir_4257, amir_4258, amir_4259. And Nostoc commune with the gene names mysA, mysB mysC and mysD. iGEM distribution kit arrived. Ordered synthetic DNA to initiate the lab work process. Ordered primers for biobricks. Primers arrived. Started the first cloning process Biobricks with PCR, followed by restriction and overnight ligation. First bacterial transformation. Resulting colonies on most of the plates. Obtained mysA biobrick sent for sequence determination. Positive result. Interlab: we took two colonies from each of transofrmation plates and inoculated them in seperate tubes of 5 mL of LB+CAM. Team photoshoot. CPEC, gybson, vegas cloning initiation. PCR with Phusion Polymerase. -Skype conference with Ecuador team Successfully completed interlab. �Achievement- Fulfilled the bronze metal requirement. Primers for gene one by one insertion, After having realized that instead of constructing a complex of wished genes and insert it to another does notlead to anything, We started to approach our goal by changing our mechanism and switching to one by one insertion of genes. Started using the blue-white screening technique while transforming samples to the plates. Lab technician day.Prepared LB media, different volume tips for autoclaving. No colonies present of p8, p9, p10. Restared with digestion of those 3 for 3 days with ECORi PSTI restrictases. p37, p39, p41. LIac mediated yeast transformation. First transformation into the yeast. Successful. last transformation to the yeast. Characterization of the biobrick BBa_K2576009 (pRPL18B promoter). Strains used:
H2O
31.6
5xHF buffer
10
25mM dNTP's (,,Template DNA)
0.4
Forward primer
2.5
Reverse primer
2.5
Template DNA
1
DM50
1.5
Phusion polymerase
0.5
