Difference between revisions of "Team:Brasil-USP/Notebook/Calendar"
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Revision as of 01:29, 18 September 2015
Calendar
Notebook
July
- PCR lasR (BBa_C0079). The purpose of this PCR was to add the RBS (BBa_K143021), through addition of the sequence in the primer foward, and also to remove the LVA tag. In our primers we only added the restriction sites X and S, so we still need to put it in the pSB1C3 for the Biobrick standard completion. When standardized this part will be called BBa_K1521001.
- Gel Purification of lasR PCR product
- Transformation of the Biobricks:
- Inoculum of the Biobricks sent by iGEM HQ:
- Transformation of the Biobricks sent by the Imperial College Team:
- Miniprep, Quantification of the plasmidial DNA samples and Restriction Analysis:
- Inoculum:
- Miniprep, Quantification and Restriction Analysis:
- BBa_K143012
- BBa_K081001
- BBa_E0840
- Transformation:
- Inoculum:
- Miniprep, Quantification and Restriction Analysis:
- BBa_B0015 (restriction analysis fail, repeat)
- PCR the qteE gene for amplification. We also added the RBS (BBa_K143021,) using the foward primer. In our primers we only added the restriction sites X and S, so we still need to put it in the pSB1C3 for the Biobrick standard completion. When standardized this biobrick will be called BBa_K1521000.
- Ligation of RBS+lasR in PTZ57R/T vector (instaclone kit)
- Prepare 40% glycerol solution
- Transformation:
- RBS + lasR (BBa_C0079)
- Gel Purification:
- RBS + qteE
- Assembly:AIII
- Prepare X-gal
- Prepare more LB medium (solid + liquid)
- Assembly:AIII
- Quantification of digestion EXSP products
- Replate RBS+lasR
- Ligation of PCR qteE in pGEM T-easy vector
- Transformation:
- Restriction analysis:
- Transformation:
- BBa_K823003
- RBS+qteE
- Electrophoresis analysis:
- Glycerol Stock and Miniprep:
-
- BBa_J04450
- RBS+lasR
- BBa_B0015
- Digestion:
- BBa_J04450 (EP)
- BBa_J04450 (XS)
- Inoculum:
- Transformation
- Glycerol Stock and Miniprep:
- Gel Purification:
- BBa_J04450 (EP)
- BBa_J04450 (XS)
- Restriction analysis:
- Transformation:
- Inoculum:
- BBa_K143055
- RBS+qteE
- Assembly KI:
- Inoculum:
- Miniprep, Restriction analysis and Digestion:
- RBS+qteE
- Assemblies AIII and KI:
- Assemblies AIII and KI:
- Prepare more competent cells
- Prepare LB medium (solid and liquid)
- Inoculum
- Prepare RBS+lasR and RBS+qteE for sequencing
- Assemblies AIII and KI:
- Glycerol Stock
- Miniprep
- Restriction analysis
- Assembly AIII:
- Restriction analysis (+NdeI enzyme)
- OBS: This analysis was reapeted because we had problems confirming the assembly. The issue was that both the construction and the pSB1C3 vector had similar size.
- Assembly AII, AV and AVI:
- Assembly AII, AV and AVI:
- Inoculum
- PCR BBa_K143055 for RBS removal
- OBS: This was one of the parts sent by the Imperial College Team. Because of a comunication failure we assumed that this part was comming without the RBS, so we included a RBS in the parts we were going to use with this Lac promotor :)
- PCR of Plac BBa_K143055 was followed by a Gel Purification and Ligation in the PUC19 vector
August
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
|
12/08
|
13/08 | 14/08
|
15/08 | 16/08 | 17/08 |
18/08 | 19/08 | 20/08 | 21/08
|
22/08
|
23/08 | 24/08 |
25/08
|
26/08 | 27/08 | 28/08 | 29/08 | 30/08 | 31/08 |
September
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
|
19/09 | 20/09 | 21/09 |
22/09
|
23/09
|
24/09
|
25/09
|
26/09 | 27/09 | 28/09 |
29/09 | 30/09 |
October
Monday | Tuesday | Wednesday | Thursday | Friday | Saturday | Sunday |
---|---|---|---|---|---|---|
01/10 | 02/10 | 03/10 | 04/10 | 05/10 | ||
06/10 | 07/10 | 08/10 | 09/10
|
10/10 | 11/10 | 12/10 |
13/10 | 14/10 | 15/10 | 16/10 | 17/10 | 18/10 | 19/10 |
20/10 | 21/10 | 22/10 | 23/10 | 24/10 | 25/10 | 26/10 |
27/10 | 28/10 | 29/10 | 30/10 | 31/10 |
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