Team:Warsaw/Calendar-Main/10 July 2009
From 2009.igem.org
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 BBa_K177024 and BBa_K177025)
Franek
Tasks:
- Alkaline lysis of the plasmid containing BBa_B0024 terminator
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.
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
Making of RBS-cI part
Jarek
Task:
- Digestion of parts BBa_C0051 and BBa_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 of p53 coding sequence
Marcin
Comment:
Due to problem with low quality of 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 K177012 operon1_part2
Ania
Tasks:
- Competent cells were transformed with LacI C0012 taken out of the distribution 2009 Kit Plate 1 well 20.
Isolation of BioBricks from 2008 and 2009 Kit Plates
Monika
Tasks:
- Isolate plasmids containing the biobricks - continuation
- GFP coding device switched on by IPTG - BBa_I763004 from 2008 Kit Plate 1017 well G5
- promoter lambda (cI regulated) with RFP reporter BBa_I763007 from 2009 Kit Plate 1 well 15J
- GFP generator - BBa_E0840 from 2009 Kit Plate 1 well 12O
- GFP generator - BBa_J07037 from 2009 Kit Plate 1 well 12O
Results from 9 July 2009
- all transformations exept GFP coding device switched on by IPTG (BBa_I763004) were successful
Methods:
- Planting on LB medium supplemented with apropriate antibiotic, 6h incubation in 37°C
- Planting on LB-agar medium supplemented with apropriate antibiotic
- Alkaline lysis (prodedure described here)
Results
- Concentration of DNA sample was measured using NanoDrop ND-1000
DNA sample | DNA concentration in ng/µl |
---|---|
BBa_I763007 | 12.63 and 17.95 |
BBa_E0840 | 34.91 and 25.66 |
BBa_J07037 | 12.52 and 48.68 |
Comment
- Isolations where DNA concentration is under 30ng/µl must be repeated
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