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Team:Warsaw/Calendar-Main/10 July 2009
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| - | + | __NOTOC__ | |
| - | + | <h3>Team meeting</h3> | |
| - | + | __NOEDITSECTION__ | |
# presentations of work did by both groups during last week (given by Ania and Kuba) | # 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 methods used by teams starting in previous editions of iGEM to kill bacteria :) (Kamil) | ||
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Methods: | Methods: | ||
| - | * [http://aabiot.com/products/dna_purification/ | + | * [http://aabiot.com/products/dna_purification/plasmid_dna/plasmid_mini/protocol_plasmid_mini.pdf <span style="color: black;">PlasmidMini set by A&A Biotechnology</span>] was used. 2 test tubes with 5ml LB and ampicillin were first inoculated with colonies containing [http://partsregistry.org/Part:BBa_B0024 <span style="color: black;">BBa_B0024</span>] brick on [http://partsregistry.org/Part:pSB1A2 <span style="color: black;">pSB1A2</span>] 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. |
<br> | <br> | ||
Results: | Results: | ||
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<!-- TU PISZ CO CHCESZ! --> | <!-- TU PISZ CO CHCESZ! --> | ||
| + | ===<div style="text-align: center;">Making of RBS-cI part</div>=== | ||
| + | |||
<html> | <html> | ||
<h4>Jarek</h4> | <h4>Jarek</h4> | ||
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<p>Task:</p> | <p>Task:</p> | ||
<ul> | <ul> | ||
| - | <li>Digestion of parts | + | <li>Digestion of parts <a href="http://partsregistry.org/Part:BBa_C0051"><span style="color: black">BBa_C0051</a></span> and <a href="http://partsregistry.org/Part:BBa_B0032"><span style="color: black">BBa_B0032</a></span></li> |
<li>Electrophoretic separation of digested parts</li> | <li>Electrophoretic separation of digested parts</li> | ||
<li>Isolation of DNA samples from gel</li> | <li>Isolation of DNA samples from gel</li> | ||
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</html> | </html> | ||
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| - | + | __NOEDITSECTION__ | |
| - | + | <html> | |
| - | Due to | + | <h3><div style="text-align: center;">Cloning of p53 coding sequence</div></h3> |
| + | <h4>Marcin</h4> | ||
| + | <br/> | ||
| + | <p><b>Comment:</p></b> | ||
| + | <p>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.</p> | ||
| + | <br/> | ||
| + | <p>Tasks:</p> | ||
| + | <ul> | ||
| + | <li>Transformation of chemocompetent strain of E. coli by plasmid containing p53 sequence</li> | ||
| + | </ul> | ||
| + | <p>Procedure:</p> | ||
| + | <ul> | ||
| + | <li>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 <a href="https://2009.igem.org/Team:Warsaw/Calendar-Main/7_July_2009">here</a> | ||
| + | </li> | ||
| + | <li>Bacteria was plated on the medium containing kanamycin</li></ul> | ||
| - | + | </html> | |
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*Competent cells were transformed with LacI C0012 taken out of the distribution 2009 Kit Plate 1 well 20. | *Competent cells were transformed with LacI C0012 taken out of the distribution 2009 Kit Plate 1 well 20. | ||
| + | |||
| + | |||
| + | ===<div style="text-align: center;">Isolation of BioBricks from 2008 and 2009 Kit Plates</div>=== | ||
| + | |||
| + | '''Monika''' | ||
| + | |||
| + | Tasks: | ||
| + | |||
| + | *Isolate plasmids containing the biobricks - continuation | ||
| + | |||
| + | # GFP coding device switched on by IPTG - [http://partsregistry.org/Part:BBa_I763004 <span style="color: black;">BBa_I763004</span>] from 2008 Kit Plate 1017 well G5 | ||
| + | # promoter lambda (cI regulated) with RFP reporter [http://partsregistry.org/Part:BBa_I763007 <span style="color: black;">BBa_I763007</span>] from 2009 Kit Plate 1 well 15J | ||
| + | # GFP generator - [http://partsregistry.org/Part:BBa_E0840 <span style="color: black;">BBa_E0840</span>] from 2009 Kit Plate 1 well 12O | ||
| + | # GFP generator - [http://partsregistry.org/Part:BBa_J07037 <span style="color: black;">BBa_J07037</span>] from 2009 Kit Plate 1 well 12O | ||
| + | |||
| + | <br> | ||
| + | |||
| + | 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 [http://www.aabiot.com/products/dna_purification/plasmid_dna/plasmid_mini/protocol_plasmid_mini.pdf <span style="color: blue;"> here</span>]) | ||
| + | |||
| + | <br> | ||
| + | |||
| + | Results | ||
| + | *Concentration of DNA sample was measured using NanoDrop ND-1000 | ||
| + | <br> | ||
| + | {| class="wikitable" style="text-align:center; width:400px; height:90px; border-collapse: collapse; border: 1px dashed blue; margin: 1em auto 1em auto" border="1" | ||
| + | |- | ||
| + | ! DNA sample | ||
| + | ! DNA concentration in ng/µl | ||
| + | |- | ||
| + | | [http://partsregistry.org/Part:BBa_I763007 <span style="color: black;">BBa_I763007</span>] | ||
| + | | 12.63 and 17.95 | ||
| + | |- | ||
| + | | [http://partsregistry.org/Part:BBa_E0840 <span style="color: black;">BBa_E0840</span>] | ||
| + | | 34.91 and 25.66 | ||
| + | |- | ||
| + | | [http://partsregistry.org/Part:BBa_J07037 <span style="color: black;">BBa_J07037</span>] | ||
| + | | 12.52 and 48.68 | ||
| + | |} | ||
| + | <br> | ||
| + | |||
| + | Comment | ||
| + | *Isolations where DNA concentration is under 30ng/µl must be repeated | ||
<!-- do not remove this! --> | <!-- do not remove this! --> | ||
{{WarNotebookEnd}} | {{WarNotebookEnd}} | ||
Latest revision as of 21:17, 19 September 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/plasmid_dna/plasmid_mini/protocol_plasmid_mini.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
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 [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.
Isolation of BioBricks from 2008 and 2009 Kit Plates
Monika
Tasks:
- Isolate plasmids containing the biobricks - continuation
- GFP coding device switched on by IPTG - [http://partsregistry.org/Part:BBa_I763004 BBa_I763004] from 2008 Kit Plate 1017 well G5
- promoter lambda (cI regulated) with RFP reporter [http://partsregistry.org/Part:BBa_I763007 BBa_I763007] from 2009 Kit Plate 1 well 15J
- GFP generator - [http://partsregistry.org/Part:BBa_E0840 BBa_E0840] from 2009 Kit Plate 1 well 12O
- GFP generator - [http://partsregistry.org/Part:BBa_J07037 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 [http://www.aabiot.com/products/dna_purification/plasmid_dna/plasmid_mini/protocol_plasmid_mini.pdf here])
Results
- Concentration of DNA sample was measured using NanoDrop ND-1000
| DNA sample | DNA concentration in ng/µl |
|---|---|
| [http://partsregistry.org/Part:BBa_I763007 BBa_I763007] | 12.63 and 17.95 |
| [http://partsregistry.org/Part:BBa_E0840 BBa_E0840] | 34.91 and 25.66 |
| [http://partsregistry.org/Part:BBa_J07037 BBa_J07037] | 12.52 and 48.68 |
Comment
- Isolations where DNA concentration is under 30ng/µl must be repeated
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