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Team:Paris-Saclay/Project/Notebook/Week 11
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| - | | style="width: 50%;"| | + | | style="width: 50%;"|Determination of the concentration of plasmid pSB1A2 which has been linearized by HINDIII. We are expecting a band at size (2079+935) ~3kbp |
| style="width: 35%;"| [[File:Week11-4.jpg|right|280px]] | | style="width: 35%;"| [[File:Week11-4.jpg|right|280px]] | ||
|- | |- | ||
| - | | style="width: 50%;"|A new PCR of the plasmid pSB1A2 is realized to obtain more quantity of the plasmid. We are expecting a band at | + | | style="width: 50%;"|A new PCR of the plasmid pSB1A2 is realized to obtain more quantity of the plasmid. We are expecting a band at size 2079 bp. |
| style="width: 35%;"| [[File:Week11-5.jpg|right|280px]] | | style="width: 35%;"| [[File:Week11-5.jpg|right|280px]] | ||
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PCR program used: | PCR program used: | ||
[[File:Week11-6.jpg|500px]] | [[File:Week11-6.jpg|500px]] | ||
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====14th August==== | ====14th August==== | ||
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{| width="85%" | {| width="85%" | ||
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| - | | style="width: 50%;"| | + | | style="width: 50%;"|Determination of the concentration of the BBa_K274100 by electrophoresis on a gel at O.8% Agarose. We are expecting a band at size 2100bp |
| style="width: 35%;"| [[File:Week11-7.jpg|right|280px]] | | style="width: 35%;"| [[File:Week11-7.jpg|right|280px]] | ||
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**PSB1A3Amil CP + Ampicilline | **PSB1A3Amil CP + Ampicilline | ||
**PSB1C3 Amil GFP + Chloramphinicol | **PSB1C3 Amil GFP + Chloramphinicol | ||
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====15th August==== | ====15th August==== | ||
| - | + | French holiday. | |
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====16th August==== | ====16th August==== | ||
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{| width="85%" | {| width="85%" | ||
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| - | | style="width: 50%;"|Visualization of the DPNI’s digestion of the plasmid pSB1A2 by 0.8% Agarose gel electrophoresis. . We are expecting a band at | + | | style="width: 50%;"|Visualization of the DPNI’s digestion of the plasmid pSB1A2 by 0.8% Agarose gel electrophoresis. . We are expecting a band at size 2079 bp. |
| style="width: 35%;"| [[File:Week11-10.jpg|right|220px]] | | style="width: 35%;"| [[File:Week11-10.jpg|right|220px]] | ||
|} | |} | ||
| - | *Miniprep of the | + | *Miniprep of the pSB1A2 plasmid. |
*Miniprep of BBa_K274100 and BBa_K115017 | *Miniprep of BBa_K274100 and BBa_K115017 | ||
*Gibson assembly of the B construction | *Gibson assembly of the B construction | ||
Latest revision as of 00:39, 27 September 2012
Week 11
13th August
| Purification by PCR clean-up of BBa_K115017 treated with DPNI. To visualize the fragment, an electrophoresis has been made with a 2% Agarose gel. We are expecting a band at the size of 123 bp. |  |
 | New PCR of BBa_K115017 to obtain more quantity of the fragment |
PCR program used:
- Digestion by DPNI of the BBa_K115017 in order to degrade the matrix plasmid.
| Determination of the concentration of plasmid pSB1A2 which has been linearized by HINDIII. We are expecting a band at size (2079+935) ~3kbp |  |
| A new PCR of the plasmid pSB1A2 is realized to obtain more quantity of the plasmid. We are expecting a band at size 2079 bp. |  |
PCR program used:
14th August
| Determination of the concentration of the BBa_K274100 by electrophoresis on a gel at O.8% Agarose. We are expecting a band at size 2100bp |  |
- Digestion of the BBa_K274100 by DPNI to degrade the matrix plasmid.
- Stocking up cells with glycerol
- PSB1A3Amil CP + Ampicilline
- PSB1C3 Amil GFP + Chloramphinicol
15th August
French holiday.
16th August
| PCR of BBa_K274100 and visualization by 0.8% Agarose gel electrophoresis. We are expecting a band at 3400bp. |  |
PCR program used:
- Digestion of the BBa_K274100 by DPNI to degrade the matrix plasmid.
17th august
| Visualization of the DPNI’s digestion of the plasmid pSB1A2 by 0.8% Agarose gel electrophoresis. . We are expecting a band at size 2079 bp. |  |
- Miniprep of the pSB1A2 plasmid.
- Miniprep of BBa_K274100 and BBa_K115017
- Gibson assembly of the B construction
- Transformation of DH5α competent cells with the Gibson B construction. The Petri dishes are placed at 37°C.
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