Team:Brasil-SP/Notebook
From 2014.igem.org
(Difference between revisions)
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</ul> | </ul> | ||
</th> | </th> | ||
- | <th> | + | <th>03/07 |
<ul> | <ul> | ||
<li>Transformation of the Biobricks sent by the Imperial College Team:</li> | <li>Transformation of the Biobricks sent by the Imperial College Team:</li> | ||
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</ul> | </ul> | ||
</ul> | </ul> | ||
- | </ | + | </th> |
- | < | + | <th>04/07</th> |
- | < | + | <th>05/07</th> |
- | < | + | <th>06/07 |
<ul> | <ul> | ||
<li>Inoculum:</li> | <li>Inoculum:</li> | ||
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</ul> | </ul> | ||
</ul> | </ul> | ||
- | </ | + | </th> |
</tr> | </tr> | ||
<tr> | <tr> | ||
- | < | + | <th>07/07 |
<ul> | <ul> | ||
<li>Miniprep, Quantification and Restriction Analysis: | <li>Miniprep, Quantification and Restriction Analysis: | ||
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</ul> | </ul> | ||
</ul> | </ul> | ||
- | </ | + | </th> |
- | < | + | <th>08/07</th> |
- | < | + | <th>09/07 |
<ul> | <ul> | ||
<li>Inoculum:</li> | <li>Inoculum:</li> | ||
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</ul> | </ul> | ||
</ul> | </ul> | ||
- | </ | + | </th> |
- | < | + | <th>10/07 |
<ul> | <ul> | ||
<li>Miniprep, Quantification and Restriction Analysis:</li> | <li>Miniprep, Quantification and Restriction Analysis:</li> | ||
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<li>Ligation of RBS+lasR in PTZ57R/T vector (instaclone kit)</li> | <li>Ligation of RBS+lasR in PTZ57R/T vector (instaclone kit)</li> | ||
</ul> | </ul> | ||
- | </ | + | </th> |
- | < | + | <th>11/07 |
<ul> | <ul> | ||
<li>Prepare 40% glycerol solution</li> | <li>Prepare 40% glycerol solution</li> | ||
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</ul> | </ul> | ||
</th> | </th> | ||
- | < | + | <th>12/07</th> |
- | < | + | <th>13/07</th> |
</tr> <tr> | </tr> <tr> | ||
- | < | + | <th>14/07 |
<ul> | <ul> | ||
<li>Assembly:<a href="https://static.igem.org/mediawiki/2014/d/d6/AIII.pdf">AIII</a></li> | <li>Assembly:<a href="https://static.igem.org/mediawiki/2014/d/d6/AIII.pdf">AIII</a></li> | ||
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<li>Prepare more LB medium (solid + liquid)</li> | <li>Prepare more LB medium (solid + liquid)</li> | ||
</ul> | </ul> | ||
- | </ | + | </th> |
- | < | + | <th>15/07 |
<ul> | <ul> | ||
<li>Assembly:<a href="https://static.igem.org/mediawiki/2014/d/d6/AIII.pdf">AIII</a></li> | <li>Assembly:<a href="https://static.igem.org/mediawiki/2014/d/d6/AIII.pdf">AIII</a></li> | ||
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</ul> | </ul> | ||
</ul> | </ul> | ||
- | </ | + | </th> |
- | < | + | <th>16/07 |
<ul> | <ul> | ||
<li>Tranformation:</li> | <li>Tranformation:</li> | ||
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</ul> | </ul> | ||
</ul> | </ul> | ||
- | </ | + | </th> |
- | < | + | <th>17/07 |
<ul> | <ul> | ||
<li>Glycerol Stock and Miniprep:<li> | <li>Glycerol Stock and Miniprep:<li> | ||
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</ul> | </ul> | ||
</ul> | </ul> | ||
- | </ | + | </th> |
- | < | + | <th>18/07 |
<ul> | <ul> | ||
<li>Glycerol Stock and Miniprep:</li> | <li>Glycerol Stock and Miniprep:</li> | ||
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</ul> | </ul> | ||
</ul> | </ul> | ||
- | </ | + | </th> |
- | < | + | <th>19/07</th> |
- | < | + | <th>20/07</th> |
</tr> <tr> | </tr> <tr> | ||
- | < | + | <th>21/07 |
<ul> | <ul> | ||
<li>Transformation:</li> | <li>Transformation:</li> | ||
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</ul> | </ul> | ||
</ul> | </ul> | ||
- | </ | + | </th> |
- | < | + | <th>22/07 |
<ul> | <ul> | ||
<li>Inoculum:</li> | <li>Inoculum:</li> | ||
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</ul> | </ul> | ||
</ul> | </ul> | ||
- | </ | + | </th> |
- | < | + | <th>23/07 |
<ul> | <ul> | ||
<li>Assemblies <a href="https://static.igem.org/mediawiki/2014/d/d6/AIII.pdf">AIII</a> and <a href="https://static.igem.org/mediawiki/2014/9/90/KI.pdf">KI</a>:</li> | <li>Assemblies <a href="https://static.igem.org/mediawiki/2014/d/d6/AIII.pdf">AIII</a> and <a href="https://static.igem.org/mediawiki/2014/9/90/KI.pdf">KI</a>:</li> | ||
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<li>Prepare RBS+lasR and RBS+qteE for sequencing</li> | <li>Prepare RBS+lasR and RBS+qteE for sequencing</li> | ||
</ul> | </ul> | ||
- | </ | + | </th> |
<td>24/07 | <td>24/07 | ||
<ul> | <ul> | ||
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</ul> | </ul> | ||
</ul> | </ul> | ||
- | </ | + | </th> |
- | < | + | <th>25/07 |
<ul> | <ul> | ||
<li>Assemblies <a href="https://static.igem.org/mediawiki/2014/d/d6/AIII.pdf">AIII</a> and <a href="https://static.igem.org/mediawiki/2014/9/90/KI.pdf">KI</a>:</li> | <li>Assemblies <a href="https://static.igem.org/mediawiki/2014/d/d6/AIII.pdf">AIII</a> and <a href="https://static.igem.org/mediawiki/2014/9/90/KI.pdf">KI</a>:</li> | ||
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</ul> | </ul> | ||
</ul> | </ul> | ||
- | </ | + | </th> |
- | < | + | <th>26/07</th> |
- | < | + | <th>27/07</th> |
</tr> <tr> | </tr> <tr> | ||
- | < | + | <th>28/07 |
<ul> | <ul> | ||
<li>Assembly <a href="https://static.igem.org/mediawiki/2014/d/d6/AIII.pdf">AIII</a>:</li> | <li>Assembly <a href="https://static.igem.org/mediawiki/2014/d/d6/AIII.pdf">AIII</a>:</li> | ||
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</ul> | </ul> | ||
</ul> | </ul> | ||
- | </ | + | </th> |
- | < | + | <th>29/07 |
<ul> | <ul> | ||
<li>>Assembly <a href="https://static.igem.org/mediawiki/2014/3/38/AII.pdf">AII</a>, | <li>>Assembly <a href="https://static.igem.org/mediawiki/2014/3/38/AII.pdf">AII</a>, | ||
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</ul> | </ul> | ||
</ul> | </ul> | ||
- | </ | + | </th> |
- | < | + | <th>30/07 |
<ul> | <ul> | ||
<li>>Assembly <a href="https://static.igem.org/mediawiki/2014/3/38/AII.pdf">AII</a>, | <li>>Assembly <a href="https://static.igem.org/mediawiki/2014/3/38/AII.pdf">AII</a>, | ||
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</ul> | </ul> | ||
</ul> | </ul> | ||
- | </ | + | </th> |
- | < | + | <th>31/07 |
<ul> | <ul> | ||
<li>Assembly <a href="https://static.igem.org/mediawiki/2014/3/38/AII.pdf">AII</a>, | <li>Assembly <a href="https://static.igem.org/mediawiki/2014/3/38/AII.pdf">AII</a>, | ||
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<li>PCR of Plac BBa_K143055 was followed by a Gel Purification and Ligation in the PUC19 vector</li> | <li>PCR of Plac BBa_K143055 was followed by a Gel Purification and Ligation in the PUC19 vector</li> | ||
</ul> | </ul> | ||
- | </ | + | </th> |
- | < | + | <th></th> |
- | < | + | <th></th> |
- | < | + | <th></th> |
</tr> | </tr> | ||
</table> | </table> | ||
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<table style="width:100%"> | <table style="width:100%"> | ||
<tr> | <tr> | ||
- | < | + | <th>Monday</th> |
- | < | + | <th>Tuesday</th> |
- | < | + | <th>Wednesday</th> |
- | < | + | <th>Thursday</th> |
- | < | + | <th>Friday</th> |
- | < | + | <th>Saturday</th> |
- | < | + | <th>Sunday</th> |
</tr> <tr> | </tr> <tr> | ||
<td></td> | <td></td> |
Revision as of 00:05, 28 September 2014
WELCOME TO iGEM 2014!Your team has been approved and you are ready to start the iGEM season!
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Notebook | ||||||||||||
You should make use of the calendar feature on the wiki and start a lab notebook. This may be looked at by the judges to see how your work progressed throughout the summer. It is a very useful organizational tool as well. |
Lab Protocols
Competent Cell with Calcium Chloride
Materials
- Solid LB medium - 1 plate
- Liquid LB medium
- 2 250ml centrifuge tubes
- Centrifuge
- Cell sample (DH5-alpha in our case)
- Autoclave
- Inoculation loop
- Liquid nitrogen
- 37°C oven
- CaCl2:
- 60mM CaCl2
- 10mM HEPES
- 15% glicerol
- H2O to 100ml
Methods
- Risk the LB plate with the cell sample (DH5-alpha)
- Incubate the plate at 37°C overnight
- Sterilize in the Autoclave
- 100ml of liquid LB medium
- 100 ml of CaCl2
- 2 250ml centrifuge tubes
- Prepare a 6ml LB inoculum with a DH5-alpha colony and incubate at 37°C/250rpm overnight
- Add 2ml of the inoculum in 100ml of liquid LB medium. Incubate at 37°C/250rpm until the OD reaches 0,375
- Distribute the volume in 2 250ml centrifuge tubes (pre chilled) and spin at 10000rpm/7min/4°C
- Discard the supernatant and ressuspend the pellet in 10ml of CaCl2 solution
- Spin the tubes at 7000rpm/7min/4°C
- Discard the supernatant and ressuspend the pellet in 10ml of CaCl2 solution. Incubate 30min on ice
- Spin at 7000rpm/7min/4°C
- Discard the supernatant and ressuspend the pellet in 2ml of the CaCl2 solution
- Distribute the 2ml in 500ul tubes adding 50ul in each.
- Freeze the samples in liquid nitrogen
- Stock at -80°C
Day 1
Day 2
Day 3
Transformation in Escherichia coli (DH5-alpha)
Materials
- 1,5ml tube
- Styrofoam box with ice
- Agar plate with antibiotic
- Competent cells
- Plasmidial DNA
- Centrifuge
- Water bath at 42°C
- Liquid LB medium
- Shaker
Methods
- Briefly spin the competent cells and put then on ice
- Add 50ng of plasmidial DNA in a 1,5ml tube
- Add the 50ul of competent cell in the same tube
- Keep the tube on ice for 25min
- Put the tube in a 42° water bath for 2 min
- Put the tube on ice for 5 min
- Add 200ul of liquid LB
- Incubate at 250rpm/37°C/1 hour
- Plate the the solution in a Agar plate with the appropriate antibiotic
- Incubate the plate at 37°C overnight
Assemblies
Life Inside the LAB
Monday | Tuesday |
Wednesday | Thursday | Friday | Saturday | Sunday |
---|---|---|---|---|---|---|
30/06
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01/07
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02/07
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03/07
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04/07 | 05/07 | 06/07
|
07/07
|
08/07 | 09/07
|
10/07
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11/07
|
12/07 | 13/07 |
14/07
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15/07
|
16/07
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17/07
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18/07
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19/07 | 20/07 |
21/07
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22/07 | 23/07 | 24/07 | 25/07 | 26/07 | 27/07 |
28/07
|
29/07 | 30/07 | 31/07
|
Monday | Tuesday | Wednesday | Thursday | Friday | Saturday | Sunday |
---|---|---|---|---|---|---|
01/08 | 02/08 | 03/08 | ||||
04/08 | 05//08 | 06/08 | 07/08
|
08/08 | 09/08 | 10/08 |
11/08
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12/08
|
13/08 | 14/08 | 15/08 | 16/08 | 17/08 |
18/08 | 19/08 | 20/08 | 21/08 | 22/08
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23/08 | 24/08 |
25/08
|
26/08 | 27/08 | 28/08 | 29/08 | 30/08 | 31/08 |
Monday | Tuesday | Wednesday | Thursday | Friday | Saturday | Sunday |
01/09 | 02/09 | 03/09 | 04/09 | 05/09 | 06/09 | 07/09 |
08/09
|
09/09 | 10/09 | 11/09 | 12/09 | 13/09 | 14/09 |
15/09 | 16/09 | 17/09
|
18/09
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19/09 | 20/09 | 21/09 |
22/09
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23/09 | 24/09 | 25/09 | 26/09 | 27/09 | 28/09 |
29/09 | 30/09 |
Monday | Tuesday | Wednesday | Thursday | Friday | Saturday | Sunday |
30/07 | 01/08 | 02/08 | 03/08 | Points | Points | |
---|---|---|---|---|---|---|
First Name | Last Name | Points | Points | Points | Points | Points |
First Name | Last Name | Points | Points | Points | Points | Points |
First Name | Last Name | Points | Points | Points | Points | Points |
First Name | Last Name | Points | Points | Points | Points | Points |