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Team:Warsaw/Calendar-Main/10 July 2009
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| - | <center><h3> | + | <center><h3>Creating devices to test promotors in <i>E. coli</i> strains (devices <a href=http://partsregistry.org/Part:BBa_K177024><font color="black">BBa_K177024</font></a> and <a href=http://partsregistry.org/Part:BBa_K177025><font color="black">BBa_K177025</font></a>)</h3></center> |
<h4>Franek</h4> | <h4>Franek</h4> | ||
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Revision as of 23:22, 11 July 2009
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Contents |
Team meeting
- presentations of work did by both groups during last week (given by Ania and Kuba)
- presentation of methods used by teams starting in previous editions of iGEM to kill bacteria :) (Kamil)
- presentation of different methods of targeting drugs to cancer cells (Marcin)
We've been also talking about some organization issues and we've decided to move our meetings - now they will take place every Thursady at 17:00, one week at the Faculty of Biology Building, another week in the room E of the lab on Pawinskiego Street.
Creating devices to test promotors in E. coli strains (devices BBa_K177024 and BBa_K177025)
Franek
Task:
Methods:
- PlasmidMini set by A&A Biotechnology was used. 2 test tubes with 5ml LB and ampicillin were first inoculated with colonies containing BBa_B0024 brick on pSB1A2 plasmid. The cultures were incubated for 6h at 37°C. Alkaline lysis was performed on both cultures. The pellet from 5 ml of bacteria was used.
- 3 test tubes with 5ml LB and ampicillin were inoculated with colonies containing BBa_R0010 brick on pSB1A2 plasmid. The cultures were left for over night incubation at 37°C.
- 3 test tubes with 5ml LB and ampicillin were inoculated with colonies containing BBa_R0080 brick on pSB1A2 plasmid. The cultures were left for overnight incubation at 37°C.
Results:
- Concentration of DNA sample was measured using NanoDrop ND-1000.
| DNA sample | DNA concentration in ng/µl |
|---|---|
| BBa_B0024 | 33.82 |
Notes:
- Plates with BBa_I0500 transformants were empty once again. This result, together with note in part description suggest that this part is damaged.
Jarek
Task:
- Digestion of parts C0051 and B0032
- Electrophoretic separation of digested parts
- Isolation of DNA samples from gel
- Ligation of part C0051 to the vector with B0032
Methods:
- DNA samples were digesteg with 1 µl of PstI/BcuI (B0032) or PstI/XbaI (C0051) in 1xTango buffer for 3 hours.
- After digestion they were separated on 0,8% agarose gel.
- Isolation of sample from gel was performed with the A&A "Gel-out" kit.
- For ligation 4µl of ligase buffer and 2 µl of ligase were used.
Results:
Cloning the p53 coding sequence
Marcin
Comment:
Due to force problem with obtaining PCR product it was necessary to transform bacteria with plasmid containing p53 sequence and to perform all procedures once more time
Tasks:
- Transformation of chemocompetent strain of E. coli by plasmid containing p53 sequence
Procedure:
- The protocol of transformation has not been changed, except of giving 1 µl of plasmid solution to the bacteria. If you want to see detailed procedure go here
- Bacteria was plated on the medium containing kanamycin
Construction of [http://partsregistry.org/wiki/index.php?title=Part:BBa_K177012 K177012] operon1_part2
Ania
Tasks:
- Competent cells were transformed with LacI C0012 taken out of the distribution 2009 Kit Plate 1 well 20.
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