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Team:Warsaw/Calendar-Main/10 July 2009
From 2009.igem.org
(Difference between revisions)
| Line 24: | Line 24: | ||
<p>Methods:</p> | <p>Methods:</p> | ||
<ul> | <ul> | ||
| - | <li>PlasmidMini set by A&A Biotechnology was used. 2 test tubes with 5ml LB and | + | <li>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. </li> |
| - | <li>3 test tubes with 5ml LB and | + | <li>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.</li> |
| - | <li>3 test tubes with 5ml LB and | + | <li>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.</li> |
</ul> | </ul> | ||
<br> | <br> | ||
| Line 37: | Line 37: | ||
|- | |- | ||
! DNA sample | ! DNA sample | ||
| - | ! DNA concentration in ng/ | + | ! DNA concentration in ng/µl |
|- | |- | ||
| BBa_B0024 | | BBa_B0024 | ||
| Line 46: | Line 46: | ||
<p>Notes:</p> | <p>Notes:</p> | ||
<ul> | <ul> | ||
| - | <li>Plates with BBa_I0500 transformants were empty once again. This result, together with note in | + | <li>Plates with BBa_I0500 transformants were empty once again. This result, together with note in <a href=http://partsregistry.org/partsdb/get_part.cgi?part=BBa_I0500>part description</a> suggest that this part is damaged.</li> |
</ul> | </ul> | ||
</html> | </html> | ||
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<p>Task:</p> | <p>Task:</p> | ||
<ul> | <ul> | ||
| - | <li>Digestion of | + | <li>Digestion of parts C0051 and B0032</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> | ||
| - | <li>Ligation of part C0051 to the | + | <li>Ligation of part C0051 to the vector with B0032<li> |
</ul> | </ul> | ||
<br> | <br> | ||
<p>Methods:</p> | <p>Methods:</p> | ||
<ul> | <ul> | ||
| - | <li>DNA samples were digesteg with 1 | + | <li>DNA samples were digesteg with 1 µl of PstI/BcuI (B0032) or PstI/XbaI (C0051) in 1xTango buffer for 3 hours.</li> |
<li>After digestion they were separated on 0,8% agarose gel.</li> | <li>After digestion they were separated on 0,8% agarose gel.</li> | ||
| - | <li>Isolation of sample from gel was performed with A&A "Gel-out" kit.</li> | + | <li>Isolation of sample from gel was performed with the <a href=http://aabiot.com/products/dna_purification/dna_fragments/gel_out/protocol_gel_out.pdf>A&A "Gel-out" kit</a>.</li> |
| - | <li>For ligation | + | <li>For ligation 4µl of ligase buffer and 2 µl of ligase were used.<li> |
</ul> | </ul> | ||
<br> | <br> | ||
| Line 88: | Line 88: | ||
Procedure: | Procedure: | ||
| - | *The protocol of transformation has not been changed, except of giving 1 | + | *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 [https://2009.igem.org/Team:Warsaw/Calendar-Main/7_July_2009 here] |
*Bacteria was plated on the medium containing canamycin | *Bacteria was plated on the medium containing canamycin | ||
Revision as of 18:28, 11 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.
Miecznikowa team: Creating devices to test promotors in E. coli strains (devices BBa_K177024 and BBa_K177025)
Franek
Task:
- Alkaline lysis of the plasmid containing BBa_B0024
- Setting cultures with BBa_R0010 and BBa_R0080
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 canamycin
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