Team:Brasil-SP/Notebook
From 2014.igem.org
(Difference between revisions)
Line 360: | Line 360: | ||
</ul> | </ul> | ||
<li>Replate RBS+lasR</li> | <li>Replate RBS+lasR</li> | ||
- | <li>Ligation of PCR qteE in pGEM vector</li> | + | <li>Ligation of PCR qteE in pGEM T-easy vector</li> |
<li>Transformation:</li> | <li>Transformation:</li> | ||
<ul> | <ul> | ||
Line 371: | Line 371: | ||
</ul> | </ul> | ||
</th> | </th> | ||
- | <th>16/07</th> | + | <th>16/07 |
- | <th>17/07</th> | + | <ul> |
- | <th>18/07</th> | + | <li>Tranformation:</li> |
+ | <ul> | ||
+ | <li>BBa_K823003</li> | ||
+ | <li>RBS+qteE</li> | ||
+ | </ul> | ||
+ | <li>Electrophoresis analysis:</li> | ||
+ | <ul> | ||
+ | <li>BBa_B0015</li> | ||
+ | </ul> | ||
+ | </ul> | ||
+ | </th> | ||
+ | <th>17/07 | ||
+ | <ul> | ||
+ | <li>Glycerol Stock and Miniprep:<li> | ||
+ | <ul> | ||
+ | <li>BBa_J04450</li> | ||
+ | <li>RBS+lasR</li> | ||
+ | <li>BBa_B0015</li> | ||
+ | </ul> | ||
+ | <li>Digestion:</li> | ||
+ | <ul> | ||
+ | <li>BBa_J04450 (EP)</li> | ||
+ | <li>BBa_J04450 (XS)</li> | ||
+ | </ul> | ||
+ | <li>Inoculum:</li> | ||
+ | <ul> | ||
+ | <li>BBa_K823003</li> | ||
+ | </ul> | ||
+ | <li>Transformation</li> | ||
+ | <ul> | ||
+ | <li>BBa_K143055</li> | ||
+ | </ul> | ||
+ | </ul> | ||
+ | </th> | ||
+ | <th>18/07 | ||
+ | <ul> | ||
+ | <li>Glycerol Stock and Miniprep:</li> | ||
+ | <ul> | ||
+ | <li>BBa_J04450</li> | ||
+ | <li>BBa_B0015</li> | ||
+ | </ul> | ||
+ | <li>Gel Purification:</li> | ||
+ | <ul> | ||
+ | <li>BBa_J04450 (EP)</li> | ||
+ | <li>BBa_J04450 (XS)</li> | ||
+ | </ul> | ||
+ | <li>Restriction analysis:</li> | ||
+ | <ul> | ||
+ | <li>BBa_K823003</li> | ||
+ | </ul> | ||
+ | </ul> | ||
+ | </th> | ||
<th>19/07</th> | <th>19/07</th> | ||
<th>20/07</th> | <th>20/07</th> |
Revision as of 00:36, 25 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
|
03/07
|
04/07 | 05/07 | 06/07
|
07/07
|
08/07 | 09/07
|
10/07
|
11/07
|
12/07 | 13/07 |
14/07
|
15/07
|
16/07
|
17/07
|
18/07
|
19/07 | 20/07 |
21/07 | 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 |
---|---|---|---|---|---|---|
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 |
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 |
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 |