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Main Page Biology Pespective Engineering Perspective Meet the Team Team Notes Pictures!

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.:: ETH Zurich - EducatETH E. coli ::.

Team descriptions (overdue) and team photograph
Christian: I guess I am the only semiprofessional photographer of the group. I can do some group pictures etc. but for this we need some ideas... - I would like to do something special. I also made the group-pics of the Synth. Biology 3.0 conference. Some references ;-) : [http://www.fotocommunity.de/pc/pc/mypics/461397]
Raphael: What about a short movie of us?
Team rosters due (1.9.)
Martin: Does somebody know, what exactly should be done here?
Nan: A list of team members, including some basic personal info.? (e.g. nationality, background, pet peeves...? )
Katerina: Guys, to make this easier, either write things about you on your personal page on the wiki or link to a page about you. This way it'll be easy to put it all together afterwards.
Labwork (parts have to be at the registry in Boston on 26.10.)
Joe: I can be in the lab at least 2 evenings a week and some times through the weekdays.
Martin: From Monday I can work every day for the whole day. At the moment I only work for several hours...
Rico: I have my exam on Tuesday. Afterwards I can assist.
Christian: I can do the introduction of the polylinker into the vectors beside my normal labwork on the Hoenggerberg. I could also do the whole biobrick assembly if you want this (I will go on holiday from the 5.10-24.10.07)
Raphael: That's the part where I will mainly contribute, from 14.09. on I can work several days/week
Christos: I can assist at the afternoons, if needed.
Katerina: 7.09 - 30.09 generally plenty of time, apart from when I do my semester project presentation, will keep you posted when that is.
Sylke: I'll not be available until 14th of October but from then on I can work several days per week fulltime.
Testing/ Analysis/ Detection (due to 02.11.)
Sylke: Meeting with Alfredo Franco-Obregon (FACS guy from center) on 18.09. concerning devices available for detection of GFP/ derivatives.
Simulation and sensitivity analysis
Martin: From 10. Sept. I've got plenty time to work on it. I think Markus would join here too ;-)
Tim: I can contribute in running stuff and help identify parameters from literature
Rico: I can do simulations, sensitivity analysis.
Nan: I would like to do simulations and sensitivity analysis.
Christos: Yeap, I guess I can be here too. I will check some toolboxes to automate things, this weekend.
Katerina: Want to help (parameter identification, programming, sensitivity analysis). Discuss in upcoming meeting tasks, versioning and ask Christian about parameters.
Presentation
Joe: I'm American... I can sale anything.
Martin: I'm bad in Layout stuff, but maybe I can help as an idea supplier or so.
Rico: I like giving presentations. For preparation we will need a mixture of different background and excellent pictures!!!
Christian: I can provide the molecular biology knowledge and part.
Nan: I can work on the slides.
Christos: I like this part, I guess everyone will contribute anyway...
Katerina: I believe I can be of help in structure, layout and fancy stuff. Could help train a bit the people we decide to do the presentation (question answering, style). I also think that Joe and Christos could be good for doing the presentation (structured, pleasant voices and lively), can discuss this in a meeting.
Sylke: I can help with the bio-part and I'm good at doing graphics and stuff. Can be a help when it needs to look good :-)
Poster
Joe: See 5. above
Rico: I can help.
Christian: I can provide the molecular biology knowledge and part.
Nan: Partly art. I will help.
Christos: You can sell, but can you trick? lol :)
Katerina: Not my strong point, but have an eye for typos, fonts, layout etc, so can help in final checking.
Sylke: See above. Good at layout.
Wiki (Project and part documentation due on 26.10)
Martin: See the points above, from next Monday I will give everything, now I'm doing my best...
Rico: I can help. I guess this will have to contain the materials that we will also use for poster and presenttion anyway.
Nan: Shouldn't it be updated with every going on process?
Christos: I am trying to put stuff in, as it comes along. I will update the bio pages with the presentation material, this weekend (I hope).
Katerina: You guys have done a great job so far, will help with whatever needed.
T-Shirts
Sylke: in print (2007-09-24)

.:: Lab Log ::.

The following are the numbering of the parts/plasmids that we use in the lab. Please use the same numberings if you label something when working in the lab. Extend the list for new concatenated parts.

[http://partsregistry.org/Part:BBa_B0034 B0034]
[http://partsregistry.org/Part:BBa_R0062 R0062]
[http://partsregistry.org/Part:BBa_R0053 R0053]
[http://partsregistry.org/Part:BBa_J23100 J23100]
[http://partsregistry.org/Part:BBa_J37033 J37033]
[http://partsregistry.org/Part:BBa_E0434 E0434]
[http://partsregistry.org/Part:BBa_B0015 B0015]
[http://partsregistry.org/Part:BBa_Q04400 Q04400]
[http://partsregistry.org/Part:BBa_R0010 R0010]
[http://partsregistry.org/Part:BBa_E0422 E0422]
[http://partsregistry.org/Part:BBa_R0040 R0040]
[http://partsregistry.org/Part:BBa_R0051 R0051]
[http://partsregistry.org/Part:BBa_Q04121 Q04121]
[http://partsregistry.org/Part:BBa_C0053 C0053]
[http://partsregistry.org/Part:BBa_Q04510 Q4510]

.:: Week 1 ::.

Date	Completed	People
Mon, 06. Aug. 2007	Preparing the Solutions	Sylke, Raphael, Stefan, Markus, Martin, Christos, Joe
Tue, 07. Aug. 2007	Prepare competent cells for all parts Transformation of all the parts	Sylke, Raphael, Stefan, Markus, Martin, Christos, Joe
Wed, 08. Aug. 2007	Preparing the grown cultures (12) for the MINIPREP (o/n cultures)	Raphael, Stefan
Thu, 09. Aug. 2007	MINIPREP of the grown (10) o/n cultures Gelelectrophoresis of the grown cultures (step: 0.8% Agarose)	Raphael, Stefan, Martin, Christos, Joe
Fri, 10. Aug. 2007	7 working parts/plasmids (step after "DIGESTS"): (B0034, R0062, R0053, E0434, B0015, R0010, E0422) 4 parts/plasmids minipreped: (R0040, R0051, Q04121, C0053)	Christos Markus Stefan
Sat, 11. Aug. 2007	no labwork
Sun, 12. Aug. 2007	labwork cancelled

.:: Week 2 ::.

Date	TODO's	Completed	People
Mon, 13. Aug. 2007 start at 3 pm	Prepare competent cells Transformations of J23100, J37033, Q04400, Q04510 Control Restrictions (step after "MINIPREP") R0040, R0051, Q04121, C0053	o/n culture (E.Coli Top10) Control Restrictions (didn't work)	Martin Markus Christos Tim
Tue, 14. Aug. 2007	Morning Shift: Start Preparing competent cells (for J23100, J37033, Q04400, Q04510) Evening Shift: Transformations of J23100, J37033, Q04400, Q04510	Morning Shift: Prepared competent cells (stored in the -80°C freezer in the basement) Evening Shift: Transformation of J23100, J37033, Q04400, Q04510 and R0040, R0051, Q04121, C0053 (in the 37°C incubator until Wednesday) Prepared new Liquid LB, LB Agar (both in the autoclave), Agarose Gel with concentrations of 0.8% and 2.4%	Morning Shift (9am-1pm?): Markus, Tim Evening Shift (5pm-...): Martin, Christos
Wed, 15. Aug. 2007	Ligation (step: "LINK ASSEMBLY"): R0053 + E0422 R0010 + E0422 R0010 + E0434 S/P: R0053, R0010 X/P: E0422, E0434	Ligation didn't work due to bad quality of enzymes (probably)	From 12: Martin, Markus
Thu, 16. Aug. 2007	Miniprep (J23100, J37033, Q04400, Q04510, R0040, R0051, Q04121, C0053) Transformation of #13 and #14	Miniprep of #4 (J23100), #5 (J37033), #8 (Q04400), #11 (R0040), #12 (R0051), #15 (Q04510) One batch is miniprepped (after step 19 in the miniprep procedure) and a second batch is frozen as a backup (which is to be miniprepped from step 3 on) Transformation of #13 (Q04121) and #14 (C0053) Numbers #13 and #14 are now growing in the 37°C incubator (step 13 in the transformation procedure)	Markus, Christos, (Martin)
Fri, 17. Aug. 2007	o/n of #13 and #14 Check whether miniprep of parts #4 #5 #8 #11 #12 (#13 #14) #15 was successful	#13 and #14 didn't grow # 4, 8 and 11 had the plasmid, they were streaked out new on plates, that we have them now on plates New white pipette tips prepared (autoclave) New bottles of Liquid LB and LB Agar prepared (autoclave)	Martin
Sat, 18. Aug. 2007
Sun, 19. Aug. 2007

.:: Week 3 ::.

Little rearrangements of the parts. Planning of the sequences to order them.

.:: Week 4 ::.

Date	TODO's	Completed	People
Mon, 27. Aug. 2007
Tue, 28. Aug. 2007
Wed, 29. Aug. 2007
Thu, 30. Aug. 2007
Fri, 31. Aug. 2007
Sat, 01. Sept. 2007	Transform pbr322, pcyc177 and pck01	Transform pbr322, pcyc177 and pck01 and plated them	Stefan
Sun, 02. Sept. 2007	Prepare o/n of pbr322, pcyc177, pck01	o/n of pcyc177, pck01 the plates of pcyc177 and pck01 are in the fridge transformed pbr322 because the culture didn't grow on the plate	Stefan

.:: Week 5 ::.

Date	TODO's	Completed	People
Mon, 03. Sept. 2007	Prepare new competent cells Miniprep pcyc177 and pcK01 prepare new o/n culture of pbr322 Run agarose gel of Minipreped plasmids	New competent cells prepared, they are now in the -80° Frezzer in the basement, column #17, dark orange box (we have now 30-35 EDTs of competent cells...) Minipreped pcyc177 and pck01 (in the -18° freezer, where the antibiotics are) pbr322 didn't grow again, so no o/n could be prepared, but we get a culture from Andy on tuesday new o/n of pcyc177 and pck01 prepared (3 Falcons each), because we need to have more plasmids 2 boxes of blue pipette tips are in the autoclave Stefan ran the agarose gel (?)	Martin, Stefan
Tue, 04. Sept. 2007	Miniprep pcyc177 and pck01 cut the prepped plasmids to test if we've got the right ones run agarose gel to test the cut and uncut ones prepare new o/n of pbr322 (from Andy)	Miniprep of pcyc177 and pck01 (but not yet tested) Prepared 3 o/ns of pbr322 (finally ;-) and each 1 o/n of pcyc177 and pck01, just in case there are problems with the miniprep	Martin, Christian
Wed, 05. Sept. 2007	Miniprep of pbr322 Test-Digest of pcyc177 and pck01 and agarose gel... Streak out all three plasmids on new plates, so we have them in reserve	New Plate of pbr322. Minipreps and Agarose Gels will be done tomorrow	Martin
Thu, 06. Sept. 2007	Miniprep of pbr322, pacyc177, pck01 Test with agarose gel	Gel of the older plasmids -> plasmid present	Christian
Fri, 07. Sept. 2007	Miniprep of pbr322, pacyc177, pck01	Plasmids miniprepped	Martin
Sat, 08. Sept. 2007
Sun, 09. Sept. 2007

.:: Week 6 ::.

Date	TODO's	Completed	People
Mon, 10. Sept. 2007	Miniprep pBR322 annealing of different MCSs Digest of pCK01 with BamHI+AseI digest of pACYC177 with BamHI+PstI digest of pBR322 with EcoRI+PstI all digests o/n		Christian
Tue, 11. Sept. 2007	Gelextraction of backbones pBR322, pCK01, pACYC digest did NOT work 1x ligation of MCS inside backbones o/d, Trafo 1x ligation of MCS inside backbones o/n	plate all 3 plasmids for new minipreps	Christian
Wed, 12. Sept. 2007	Trafo of o/n ligations o/n cultures of putative clones		Christian
Thu, 13. Sept. 2007	Minipreps of putative clones pCK01-MCS and pBR322-MCS control digests of putative clones new o/n cultures of the putative clones of o/n ligations		Christian
Fri, 14. Sept. 2007	separation of control digests of putative clones	*pBR322-MCS (Tet-selection) clone2 positive	Christian
Sat, 15. Sept. 2007
Sun, 16. Sept. 2007

.:: Week 7 ::.

Date	TODO's	Completed	People
Mon, 17. Sept. 2007	Hönggerberg: new digest of pACYC177 with BamHI+PstI o/n Zentrum: digest of pACYC177, pBR322 AP ligation of 177 and 322AP	Zentrum: digest of pBR322 AP (the concentration of DNA was too low for pacyc177...) ligation of pBR322 o/n 100 ml o/n culture to MAXIprep pacyc177 Transformation of pBR322 AP to have it on plates (because andy only miniprepped them)	Hönngerberg: Christian Zentrum: Martin, Raphael
Tue, 18. Sept. 2007	Hönggerberg: different control digests of pBR322-MCS (Tet) (see last week) separation of pACYC177 digest Zentrum: Test Digests of pck01 with XbaI, SpeI, PstI, Xba/Pst, Xba/Spe (because all of them should be in the plasmid due to the sequence, and if they are it would be crap!!!) Transformation of the ligated pbr322 AP (MCS) Prep pacyc177 Digest prepped pacyc177	Hönggerberg: > no DNA on pACYC177 digest-gel, only degradation smear Zentrum: Plates of pbr322 AP grew No Digest of pck01 worked due to too low DNA concentration... (che cazzo di low copy plasmids !!!!) Miniprepped only 20 ml of the pacyc o/n culture with Quiagen Kit, the results were great! We have loads of DNA! (thank god! ) Digest of pacyc177 with BamHI (45 µl), then precipitated, in the gel was still very much DNA, but there were still 3 bands, so we guess, that it hasn't cut, maybe because the BamHI in the center is very old, perhaps we should Digest it in Höngg again. Digest of pacyc177 with PstI o/n (pray that it will work!) New o/n cultures of pck01 (to prep it like pacyc177), pbr322 AP (to prep it too, to have something on stock again, if the ligation didn't work), top10 (to make new competent cells) test digest of pck01 with notI, but due to the low DNA concentration I don't think it will work. I took glooves, if it now work, then we have caught some DNases in the earlier test digests	Höngg: Christian Zentrum: Martin, Raphael
Wed, 19. Sept. 2007	Zentrum: o/n culture of pbr322 AP (MCS), then test digest and see if it is ligated Prep of pck01 and test digests (xba, pst, spe, pvuI, notI) check the digests of pacyc177 (pst) and pck01 (notI) design new linkers for pck01, design primers for PCR for the extraction of SpeI from pck01
Thu, 20. Sept. 2007
Fri, 21. Sept. 2007
Sat, 22. Sept. 2007
Sun, 23. Sept. 2007

.:: Week 8 ::.

Date	TODO's	Completed	People
Mon, 24. Sept. 2007
Tue, 25. Sept. 2007
Wed, 26. Sept. 2007
Thu, 27. Sept. 2007
Fri, 28. Sept. 2007
Sat, 29. Sept. 2007
Sun, 30. Sept. 2007

**.::Plasmid 1 (pbr322ap)::.**

Put parts 1,2,3 in pbr322ap plasmids.
Merge plasmid containing part 2 (donor) with plasmid containing part 3 (acceptor). You should get a plasmid containing a 2.3 composite part.
Merge plasmid containing part 1 (donor) with plasmid containing composite part 2.3 (acceptor). You should get a plasmid containing a 1.2.3 composite part.

**.::Plasmid 2 (pck01cm)::.**

Put parts 4,5,8,9 in pck01cm plasmids.
Merge plasmid containing part 4 (donor) with plasmid containing part 5 (acceptor). You should get a plasmid containing a 4.5 composite part.
Merge plasmid containing part 8 (donor) with plasmid containing part 9 (acceptor). You should get a plasmid containing a 8.9 composite part. Note: this step can be done simultaneously with the above.
Merge plasmid containing composite part 4.5 (donor) with plasmid containing composite part 8.9 (acceptor). You should get a plasmid containing a 4.5.8.9 composite part.

'.::Plasmid 3 (pacyc177km)::.

Put parts 6,7,10,11 in pacyc177km plasmids.
Merge plasmid containing part 6 (donor) with plasmid containing part 7 (acceptor). You should get a plasmid containing a 6.7 composite part.
Merge plasmid containing part 10 (donor) with plasmid containing part 11 (acceptor). You should get a plasmid containing a 10.11 composite part. Note: this step can be done simultaneously with the above.
Merge plasmid containing composite part 6.7 (donor) with plasmid containing composite part 10.11 (acceptor). You should get a plasmid containing a 6.7.10.11 composite part.

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