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.
Date
| TODO's
| Completed
| People
|
Mon, 06. Aug. 2007
|
|
| Sylke, Raphael, Stefan, Markus, Martin, Christos, Joe
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Tue, 07. Aug. 2007
|
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- Prepare competent cells for all parts
- Transformation of all the parts
| Sylke, Raphael, Stefan, Markus, Martin, Christos, Joe
|
Wed, 08. Aug. 2007
|
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- Preparing the grown cultures (12) for the MINIPREP
(o/n cultures)
| Raphael, Stefan
|
Thu, 09. Aug. 2007
|
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- 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
|
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Sun, 12. Aug. 2007
|
| labwork cancelled
|
|
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
|
|
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Sun, 19. Aug. 2007
|
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Little rearrangements of the parts. Planning of the sequences to order them.
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
|
| Martin
|
Sat, 08. Sept. 2007
|
|
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Sun, 09. Sept. 2007
|
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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
|
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Sun, 16. Sept. 2007
|
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