Team:Warsaw/Calendar-Main/10 July 2009
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<!-- TU EDYTUJE FRANEK, NIE RUSZ! --> | <!-- TU EDYTUJE FRANEK, NIE RUSZ! --> | ||
- | === | + | ===<div style="text-align: center;">Creating devices to test promoters in <em>E. coli</em> strains (devices [http://partsregistry.org/Part:BBa_K177024 <span style="color: black;">BBa_K177024</span>] and [http://partsregistry.org/Part:BBa_K177025 <span style="color: black;">BBa_K177025</span>])</div>=== |
- | ''' | + | '''Franek''' |
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Revision as of 14:56, 12 July 2009
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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 promoters in E. coli strains (devices [http://partsregistry.org/Part:BBa_K177024 BBa_K177024] and [http://partsregistry.org/Part:BBa_K177025 BBa_K177025])
Franek
Tasks:
- Alkaline lysis of the plasmid containing [http://partsregistry.org/Part:BBa_B0024 BBa_B0024 terminator]
Methods:
- [http://aabiot.com/products/dna_purification/dna_fragments/gel_out/protocol_gel_out.pdf PlasmidMini set by A&A Biotechnology] was used. 2 test tubes with 5ml LB and ampicillin were first inoculated with colonies containing [http://partsregistry.org/Part:BBa_B0024 BBa_B0024] brick on [http://partsregistry.org/Part:pSB1A2 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.
Results:
- Concentration of DNA sample was measured using NanoDrop ND-1000
DNA sample | DNA concentration in ng/µl |
---|---|
[http://partsregistry.org/Part:BBa_B0024 BBa_B0024] | 33.82 |
Notes:
- Plates with [http://partsregistry.org/Part:BBa_I0500 BBa_I0500] transformants were empty once again. This result, together with note in [http://partsregistry.org/partsdb/get_part.cgi?part=BBa_I0500 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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