iGEM 2014 — Team Brasil-SP

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# Team:Brasil-SP/Notebook

### From 2014.igem.org

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[![HeaderNotebookBRASILAP.png](/wiki/images/4/4f/HeaderNotebookBRASILAP.png)](/File%3AHeaderNotebookBRASILAP.png)

* [![](https://static.igem.org/mediawiki/2014/2/23/InicialBRASILSP.png)](https://2014.igem.org/Team%3ABrasil-SP)
* [![](https://static.igem.org/mediawiki/2014/d/df/ProblemaBRASILSP.png)](https://2014.igem.org/Team%3ABrasil-SP/TheIssue)
  + [Read more](https://2014.igem.org/Team%3ABrasil-SP/TheIssue)
* [![](https://static.igem.org/mediawiki/2014/b/bc/ProjetoBRASILSP.png)](https://2014.igem.org/Team%3ABrasil-SP/Project)
  + [Overview](https://2014.igem.org/Team%3ABrasil-SP/Project)
  + [Cystatin C](https://2014.igem.org/Team%3ABrasil-SP/Project/Cystatin)
  + [Detection Module](https://2014.igem.org/Team%3ABrasil-SP/Project/DetectionModule)
  + [Diagnosis Module](https://2014.igem.org/Team%3ABrasil-SP/Project/DiagnosisModule)
  + [Response Module](https://2014.igem.org/Team%3ABrasil-SP/Project/ResponseModule)
  + [The Device](https://2014.igem.org/Team%3ABrasil-SP/Project/Device)
* [![](https://static.igem.org/mediawiki/2014/a/af/ResultadosBRASILSP.png)](https://2014.igem.org/Team%3ABrasil-SP/Results/MainAssemblyMap)
  + [Main Assembly Map](https://2014.igem.org/Team%3ABrasil-SP/Results/MainAssemblyMap)
  + [Characterization Assemblies](https://2014.igem.org/Team%3ABrasil-SP/Results/CharacterizationAssemblies)
  + [Sporulation and Germination](https://2014.igem.org/Team%3ABrasil-SP/Results/Sporulation)
  + [Interlab study](https://2014.igem.org/Team%3ABrasil-SP/Results/interlabstudy)
  + [Biobricks](https://2014.igem.org/Team%3ABrasil-SP/Parts)
* [![](https://static.igem.org/mediawiki/2014/e/e5/ModelagemBRASILSP.png)](https://2014.igem.org/Team%3ABrasil-SP/Modeling/Detectionmodule)
  + [Detection, Diagnosis and Response Module](https://2014.igem.org/Team%3ABrasil-SP/Modeling/Detectionmodule)
* [![](https://static.igem.org/mediawiki/2014/3/3b/CadernodeLabBRASILSP.png)](https://2014.igem.org/Team%3ABrasil-SP/Notebook)
  + [Lab Journal](https://2014.igem.org/Team%3ABrasil-SP/Notebook)
  + [Protocols](https://2014.igem.org/Team%3ABrasil-SP/Notebook/Protocols)
  + [Lab Safety](https://2014.igem.org/Team%3ABrasil-SP/Safety)
* [![](https://static.igem.org/mediawiki/2014/4/43/OutreachBRASILSP.png)](https://2014.igem.org/Team%3ABrasil-SP/Outreach/BioEtics)
  + [Public Opinion on SynBio and Bioethics](https://2014.igem.org/Team%3ABrasil-SP/Outreach/BioEtics)
  + [Jamboré - The Brazilian Jamboree](https://2014.igem.org/Team%3ABrasil-SP/Outreach/Jambor%C3%A9)
  + [Working Together](https://2014.igem.org/Team%3ABrasil-SP/Outreach/WorkingTogether)
  + [Society within the Academy](https://2014.igem.org/Team%3ABrasil-SP/Outreach/Society_within_the_Academy)
  + [Media Coverage](https://2014.igem.org/Team%3ABrasil-sp/media_coverage)
* [![](https://static.igem.org/mediawiki/2014/8/81/TimeBRASILSP.png)](https://2014.igem.org/Team%3ABrasil-SP/Team)
  + [Team Members](https://2014.igem.org/Team%3ABrasil-SP/Team)
  + [Sponsors and Support](https://2014.igem.org/Team%3ABrasil-SP/Team/Sponsors)
  + [Team Official Profile](https://igem.org/Team.cgi?id=1521)
  + [Achievements](https://2014.igem.org/Team%3ABrasil-SP/Achievements)
* [![](https://static.igem.org/mediawiki/igem.org/6/60/Igemlogo_300px.png)](https://2014.igem.org/Main_Page)

# Main Assembly Map

The main assembly map describes all the constructions that would be done in the project, but some of them have not been performed in time of the iGEM deadline. The succesfully constructed assemblies is indicated by a green check point and the unsuccessfull constructions are indicated by a red "X". Click on these assemblies to access the lab form.

[AI](https://static.igem.org/mediawiki/2014/b/b7/AI.pdf)
[AII](https://static.igem.org/mediawiki/2014/3/38/AII.pdf)
[AIII](https://static.igem.org/mediawiki/2014/d/d6/AIII.pdf)
[BIII](https://static.igem.org/mediawiki/2014/e/ea/BIII.pdf)
[AIV](https://static.igem.org/mediawiki/2014/e/eb/AIV.pdf)
[BII](https://static.igem.org/mediawiki/2014/4/48/BII.pdf)
[CII](https://static.igem.org/mediawiki/2014/a/a3/CII.pdf)
[DI](https://static.igem.org/mediawiki/2014/7/72/DI.pdf)
[AVI](https://static.igem.org/mediawiki/2014/d/d6/AVI.pdf)
[AV](https://static.igem.org/mediawiki/2014/f/fe/AV.pdf)
[BIV](https://static.igem.org/mediawiki/2014/5/5b/BIV.pdf)

# Assemblies forms

**Assembly form template**

This is the template designed to help with the laboratory organization during the assemble of biological parts. If you want to use the assembly method we used you can print the form and complete it for each construction. Hope it helps :)

[![](https://static.igem.org/mediawiki/2014/3/32/Assembly_form_pg3.png)](https://static.igem.org/mediawiki/2014/e/eb/Assembly_Form.pdf)

# **Life Inside the LAB**

## July

| Monday | Tuesday | Wednesday | Thursday | Friday | Saturday | Sunday |
| --- | --- | --- | --- | --- | --- | --- |
| 30/06  * [PCR](https://static.igem.org/mediawiki/2014/7/74/PCR.pdf) lasR ([BBa\_C0079](http://parts.igem.org/Part%3ABBa_C0079)). The purpose of this PCR was to add the RBS ([BBa\_K143021](http://parts.igem.org/Part%3ABBa_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](http://parts.igem.org/Part%3ABBa_K1521001). | 01/07  * [Gel Purification](https://static.igem.org/mediawiki/2014/a/af/Agarose_Gel_Electrophoresis_and_DNA_Gel_Purification.pdf) of lasR [PCR](https://static.igem.org/mediawiki/2014/7/74/PCR.pdf) product | 02/07  * [Transformation](https://static.igem.org/mediawiki/2014/a/a5/Transformation_in_Escherichia_coli_DH5.pdf) of the Biobricks:  + [BBa\_K143012](http://parts.igem.org/Part%3ABBa_K143012) + [BBa\_K081001](http://parts.igem.org/Part%3ABBa_K081001) + [BBa\_E0840](http://parts.igem.org/Part%3ABBa_E0840)  * Inoculum of the Biobricks sent by iGEM HQ:  + [BBa\_K316037](http://parts.igem.org/Part%3ABBa_K316037) + [BBa\_K316018](http://parts.igem.org/Part%3ABBa_K316018) + [BBa\_K316015](http://parts.igem.org/Part%3ABBa_K316015) | 03/07  * [Transformation](https://static.igem.org/mediawiki/2014/a/a5/Transformation_in_Escherichia_coli_DH5.pdf) of the Biobricks sent by the Imperial College Team:  + [BBa\_K316016](http://parts.igem.org/Part%3ABBa_K316016) + [BBa\_K143031](http://parts.igem.org/Part%3ABBa_K143031)  * [Miniprep](https://static.igem.org/mediawiki/2014/6/6f/Purelink_quick_plasmid_qrc.pdf), Quantification of the plasmidial DNA samples and [Restriction Analysis](https://static.igem.org/mediawiki/2014/4/42/Restriction_Anaysis_and_Digestion_in_large_scale.pdf):  + [BBa\_K316037](http://parts.igem.org/Part%3ABBa_K316037) + [BBa\_K316018](http://parts.igem.org/Part%3ABBa_K316018) + [BBa\_K316015](http://parts.igem.org/Part%3ABBa_K316015) | 04/07 | 05/07 | 06/07  * Inoculum:  + [BBa\_K143012](http://parts.igem.org/Part%3ABBa_K143012) + [BBa\_K081001](http://parts.igem.org/Part%3ABBa_K081001) + [BBa\_E0840](http://parts.igem.org/Part%3ABBa_E0840) |
| 07/07  * [Miniprep](https://static.igem.org/mediawiki/2014/6/6f/Purelink_quick_plasmid_qrc.pdf), Quantification and [Restriction Analysis](https://static.igem.org/mediawiki/2014/4/42/Restriction_Anaysis_and_Digestion_in_large_scale.pdf):  + [BBa\_K143012](http://parts.igem.org/Part%3ABBa_K143012) + [BBa\_K081001](http://parts.igem.org/Part%3ABBa_K081001) BBa\_E0840  * [Transformation](https://static.igem.org/mediawiki/2014/a/a5/Transformation_in_Escherichia_coli_DH5.pdf):  + BBa\_B0015 | 08/07 | 09/07  * Inoculum:  + BBa\_B0015 | 10/07  * [Miniprep](https://static.igem.org/mediawiki/2014/6/6f/Purelink_quick_plasmid_qrc.pdf), Quantification and [Restriction Analysis](https://static.igem.org/mediawiki/2014/4/42/Restriction_Anaysis_and_Digestion_in_large_scale.pdf):  + BBa\_B0015 (restriction analysis fail, repeat)  * [PCR](https://static.igem.org/mediawiki/2014/7/74/PCR.pdf) the qteE gene for amplification. We also added the RBS ([BBa\_K143021,](http://parts.igem.org/Part%3ABBa_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](http://parts.igem.org/Part%3ABBa_K1521000). * [Ligation](https://static.igem.org/mediawiki/2014/0/03/PCR_Product_Clonig.pdf) of RBS+lasR in PTZ57R/T vector (instaclone kit) | 11/07  * Prepare 40% glycerol solution * [Transformation](https://static.igem.org/mediawiki/2014/a/a5/Transformation_in_Escherichia_coli_DH5.pdf):  + RBS + lasR ([BBa\_C0079](http://parts.igem.org/Part%3ABBa_C0079))  * [Gel Purification](https://static.igem.org/mediawiki/2014/a/af/Agarose_Gel_Electrophoresis_and_DNA_Gel_Purification.pdf):  + RBS + qteE | 12/07 | 13/07 |
| 14/07  * Assembly:[AIII](https://static.igem.org/mediawiki/2014/d/d6/AIII.pdf)  + [Digestion EXSP](https://static.igem.org/mediawiki/2014/4/42/Restriction_Anaysis_and_Digestion_in_large_scale.pdf)  * Prepare X-gal * Prepare more LB medium (solid + liquid) | 15/07  * Assembly:[AIII](https://static.igem.org/mediawiki/2014/d/d6/AIII.pdf)  + Quantification of digestion EXSP products  * Replate RBS+lasR * [Ligation](https://static.igem.org/mediawiki/2014/0/03/PCR_Product_Clonig.pdf) of [PCR](https://static.igem.org/mediawiki/2014/7/74/PCR.pdf) qteE in pGEM T-easy vector * [Transformation](https://static.igem.org/mediawiki/2014/a/a5/Transformation_in_Escherichia_coli_DH5.pdf):  + BBa\_J04450  * [Restriction analysis](https://static.igem.org/mediawiki/2014/4/42/Restriction_Anaysis_and_Digestion_in_large_scale.pdf):  + BBa\_B0015 | 16/07  * [Transformation](https://static.igem.org/mediawiki/2014/a/a5/Transformation_in_Escherichia_coli_DH5.pdf):  + [BBa\_K823003](http://parts.igem.org/Part%3ABBa_K823003) + RBS+qteE  * Electrophoresis analysis:  + BBa\_B0015 | 17/07  * Glycerol Stock and [Miniprep](https://static.igem.org/mediawiki/2014/6/6f/Purelink_quick_plasmid_qrc.pdf):* + [BBa\_J04450](http://parts.igem.org/Part%3ABBa_J04450)     + RBS+lasR     + BBa\_B0015* [Digestion](https://static.igem.org/mediawiki/2014/4/42/Restriction_Anaysis_and_Digestion_in_large_scale.pdf):     + [BBa\_J04450](http://parts.igem.org/Part%3ABBa_J04450) (EP)     + [BBa\_J04450](http://parts.igem.org/Part%3ABBa_J04450) (XS)* Inoculum:     + [BBa\_K823003](http://parts.igem.org/Part%3ABBa_K823003)* [Transformation](https://static.igem.org/mediawiki/2014/a/a5/Transformation_in_Escherichia_coli_DH5.pdf)     + [BBa\_K143055](http://parts.igem.org/Part%3ABBa_K143055) | 18/07  * Glycerol Stock and [Miniprep](https://static.igem.org/mediawiki/2014/6/6f/Purelink_quick_plasmid_qrc.pdf):  + [BBa\_J04450](http://parts.igem.org/Part%3ABBa_J04450) + BBa\_B0015  * [Gel Purification](https://static.igem.org/mediawiki/2014/a/af/Agarose_Gel_Electrophoresis_and_DNA_Gel_Purification.pdf):  + [BBa\_J04450](http://parts.igem.org/Part%3ABBa_J04450) (EP) + [BBa\_J04450](http://parts.igem.org/Part%3ABBa_J04450) (XS)  * [Restriction analysis](https://static.igem.org/mediawiki/2014/4/42/Restriction_Anaysis_and_Digestion_in_large_scale.pdf):  + [BBa\_K823003](http://parts.igem.org/Part%3ABBa_K823003) | 19/07 | 20/07 |
| 21/07  * [Transformation](https://static.igem.org/mediawiki/2014/a/a5/Transformation_in_Escherichia_coli_DH5.pdf):  + [BBa\_K316016](http://parts.igem.org/Part%3ABBa_K316016)  * Inoculum:  + [BBa\_K143055](http://parts.igem.org/Part%3ABBa_K143055) + RBS+qteE  * Assembly [KI](https://static.igem.org/mediawiki/2014/9/90/KI.pdf):  + [Digestion EXSP](https://static.igem.org/mediawiki/2014/4/42/Restriction_Anaysis_and_Digestion_in_large_scale.pdf) | 22/07  * Inoculum:  + [BBa\_K316016](http://parts.igem.org/Part%3ABBa_K316016)  * [Miniprep](https://static.igem.org/mediawiki/2014/6/6f/Purelink_quick_plasmid_qrc.pdf), [Restriction analysis](https://static.igem.org/mediawiki/2014/4/42/Restriction_Anaysis_and_Digestion_in_large_scale.pdf) and [Digestion](https://static.igem.org/mediawiki/2014/4/42/Restriction_Anaysis_and_Digestion_in_large_scale.pdf):  + RBS+qteE  * Assemblies [AIII](https://static.igem.org/mediawiki/2014/d/d6/AIII.pdf) and [KI](https://static.igem.org/mediawiki/2014/9/90/KI.pdf):  + [Gel Purification](https://static.igem.org/mediawiki/2014/a/af/Agarose_Gel_Electrophoresis_and_DNA_Gel_Purification.pdf) + [Ligation](https://static.igem.org/mediawiki/2014/7/7a/Cohesive-end_assembly_cloning.pdf) | 23/07  * Assemblies [AIII](https://static.igem.org/mediawiki/2014/d/d6/AIII.pdf) and [KI](https://static.igem.org/mediawiki/2014/9/90/KI.pdf):  + [Transformation](https://static.igem.org/mediawiki/2014/a/a5/Transformation_in_Escherichia_coli_DH5.pdf)  * Prepare more  [competent cells](https://static.igem.org/mediawiki/2014/9/96/Competent_Cells_using_Calcium_Chloride.pdf) * Prepare LB medium (solid and liquid) * Inoculum  + [BBa\_K316016](http://parts.igem.org/Part%3ABBa_K316016)  * Prepare RBS+lasR and RBS+qteE for sequencing | 24/07  * Assemblies [AIII](https://static.igem.org/mediawiki/2014/d/d6/AIII.pdf) and [KI](https://static.igem.org/mediawiki/2014/9/90/KI.pdf):  + Inoculum  * [Miniprep](https://static.igem.org/mediawiki/2014/6/6f/Purelink_quick_plasmid_qrc.pdf):  + [BBa\_K316016](http://parts.igem.org/Part%3ABBa_K316016) | 25/07  * Assemblies [AIII](https://static.igem.org/mediawiki/2014/d/d6/AIII.pdf) and [KI](https://static.igem.org/mediawiki/2014/9/90/KI.pdf):  + Glycerol Stock + [Miniprep](https://static.igem.org/mediawiki/2014/6/6f/Purelink_quick_plasmid_qrc.pdf) + [Restriction analysis](https://static.igem.org/mediawiki/2014/4/42/Restriction_Anaysis_and_Digestion_in_large_scale.pdf) | 26/07 | 27/07 |
| 28/07  * Assembly [AIII](https://static.igem.org/mediawiki/2014/d/d6/AIII.pdf):  + [Restriction analysis](https://static.igem.org/mediawiki/2014/4/42/Restriction_Anaysis_and_Digestion_in_large_scale.pdf) (+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](https://static.igem.org/mediawiki/2014/3/38/AII.pdf),   [AV](https://static.igem.org/mediawiki/2014/f/fe/AV.pdf) and   [AVI](https://static.igem.org/mediawiki/2014/d/d6/AVI.pdf):  + [Digestion EXSP](https://static.igem.org/mediawiki/2014/4/42/Restriction_Anaysis_and_Digestion_in_large_scale.pdf) | 29/07  * >Assembly [AII](https://static.igem.org/mediawiki/2014/3/38/AII.pdf),   [AV](https://static.igem.org/mediawiki/2014/f/fe/AV.pdf) and   [AVI](https://static.igem.org/mediawiki/2014/d/d6/AVI.pdf):  + [Gel Purification](https://static.igem.org/mediawiki/2014/a/af/Agarose_Gel_Electrophoresis_and_DNA_Gel_Purification.pdf) + [Ligation](https://static.igem.org/mediawiki/2014/7/7a/Cohesive-end_assembly_cloning.pdf) | 30/07  * >Assembly [AII](https://static.igem.org/mediawiki/2014/3/38/AII.pdf),   [AV](https://static.igem.org/mediawiki/2014/f/fe/AV.pdf) and   [AVI](https://static.igem.org/mediawiki/2014/d/d6/AVI.pdf):  + [Transformation](https://static.igem.org/mediawiki/2014/a/a5/Transformation_in_Escherichia_coli_DH5.pdf) | 31/07  * Assembly [AII](https://static.igem.org/mediawiki/2014/3/38/AII.pdf),   [AV](https://static.igem.org/mediawiki/2014/f/fe/AV.pdf) and   [AVI](https://static.igem.org/mediawiki/2014/d/d6/AVI.pdf):  + Inoculum  * [PCR](https://static.igem.org/mediawiki/2014/7/74/PCR.pdf) [BBa\_K143055](http://parts.igem.org/Part%3ABBa_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](http://parts.igem.org/Part%3ABBa_K143055) was followed by a [Gel Purification](https://static.igem.org/mediawiki/2014/a/af/Agarose_Gel_Electrophoresis_and_DNA_Gel_Purification.pdf) and [Ligation](https://static.igem.org/mediawiki/2014/0/03/PCR_Product_Clonig.pdf) in the PUC19 vector |  |  |  |

## August

| Monday | Tuesday | Wednesday | Thursday | Friday | Saturday | Sunday |
| --- | --- | --- | --- | --- | --- | --- |
|  |  |  |  | 01/08  * [Transformation](https://static.igem.org/mediawiki/2014/a/a5/Transformation_in_Escherichia_coli_DH5.pdf):  + [BBa\_K143055](http://parts.igem.org/Part%3ABBa_K143055)  * Assembly [AII](https://static.igem.org/mediawiki/2014/3/38/AII.pdf),   [AV](https://static.igem.org/mediawiki/2014/f/fe/AV.pdf) and   [AVI](https://static.igem.org/mediawiki/2014/d/d6/AVI.pdf):  + Glycerol Stock + [Miniprep](https://static.igem.org/mediawiki/2014/6/6f/Purelink_quick_plasmid_qrc.pdf) + [Restriction analysis](https://static.igem.org/mediawiki/2014/4/42/Restriction_Anaysis_and_Digestion_in_large_scale.pdf) | 02/08 | 03/08 |
| 04/08  * Inoculum  + E. coli DH5-alpha + [KI](https://static.igem.org/mediawiki/2014/9/90/KI.pdf) + Plac [BBa\_K143055](http://parts.igem.org/Part%3ABBa_K143055)  * [Transformation](https://static.igem.org/mediawiki/2014/a/a5/Transformation_in_Escherichia_coli_DH5.pdf)  + [BBa\_P0312](http://parts.igem.org/Part%3ABBa_P0312)  * Assembly [AV](https://static.igem.org/mediawiki/2014/f/fe/AV.pdf):  + Prepare more inoculum + Repeat [Restriction analysis](https://static.igem.org/mediawiki/2014/4/42/Restriction_Anaysis_and_Digestion_in_large_scale.pdf) | 05//08  * Prepare assemblies [KI](https://static.igem.org/mediawiki/2014/9/90/KI.pdf) for cytometry characterization * [Miniprep](https://static.igem.org/mediawiki/2014/6/6f/Purelink_quick_plasmid_qrc.pdf):  + Plac [BBa\_K143055](http://parts.igem.org/Part%3ABBa_K143055)  * Inoculum  + [BBa\_P0312](http://parts.igem.org/Part%3ABBa_P0312)  * Assembly [AV](https://static.igem.org/mediawiki/2014/f/fe/AV.pdf):  + [Miniprep](https://static.igem.org/mediawiki/2014/6/6f/Purelink_quick_plasmid_qrc.pdf) + And once again! [Restriction analysis](https://static.igem.org/mediawiki/2014/4/42/Restriction_Anaysis_and_Digestion_in_large_scale.pdf) (this [BBa\_K316016](http://parts.igem.org/Part%3ABBa_K316016) is tough) | 06/08 | 07/08  * [Miniprep](https://static.igem.org/mediawiki/2014/6/6f/Purelink_quick_plasmid_qrc.pdf) and [Restriction analysis](https://static.igem.org/mediawiki/2014/4/42/Restriction_Anaysis_and_Digestion_in_large_scale.pdf):  + [BBa\_P0312](http://parts.igem.org/Part%3ABBa_P0312) | 08/08 | 09/08 | 10/08 |
| 11/08  * Prepare Plac [BBa\_K143055](http://parts.igem.org/Part%3ABBa_K143055) for sequencing * Analyse the RBS+qteE and RBS+lasR sequencing file | 12/08  * [Digestion](https://static.igem.org/mediawiki/2014/4/42/Restriction_Anaysis_and_Digestion_in_large_scale.pdf):  + RBS+qteE + RBS+lasR + OBS: this digestion was performed so we clone this parts in the pSB1C3 vector to put them in Biobrick standard | 13/08  * Assemblies [BII](https://static.igem.org/mediawiki/2014/4/48/BII.pdf) and   [BIII](https://static.igem.org/mediawiki/2014/e/ea/BIII.pdf):  + [Digestion EXSP](https://static.igem.org/mediawiki/2014/4/42/Restriction_Anaysis_and_Digestion_in_large_scale.pdf) | 14/08  * [Gel Purification](https://static.igem.org/mediawiki/2014/a/af/Agarose_Gel_Electrophoresis_and_DNA_Gel_Purification.pdf) and [Ligation](https://static.igem.org/mediawiki/2014/7/7a/Cohesive-end_assembly_cloning.pdf) in pSB1C3 for BBiobrick standard:  + RBS+qteE + RBS+lasR  * Assemblies [BII](https://static.igem.org/mediawiki/2014/4/48/BII.pdf) and   [BIII](https://static.igem.org/mediawiki/2014/e/ea/BIII.pdf):  + [Gel Purification](https://static.igem.org/mediawiki/2014/a/af/Agarose_Gel_Electrophoresis_and_DNA_Gel_Purification.pdf) + OBS: we had some problems here with the purification and we lost all our digestions, so we're repeating the digestion | 15/08 | 16/08  * Assemblies [BII](https://static.igem.org/mediawiki/2014/4/48/BII.pdf) and   [BIII](https://static.igem.org/mediawiki/2014/e/ea/BIII.pdf):  + [Digestion EXSP](https://static.igem.org/mediawiki/2014/4/42/Restriction_Anaysis_and_Digestion_in_large_scale.pdf) | 17/08  * Assemblies [BII](https://static.igem.org/mediawiki/2014/4/48/BII.pdf) and   [BIII](https://static.igem.org/mediawiki/2014/e/ea/BIII.pdf):  + [Gel Purification](https://static.igem.org/mediawiki/2014/a/af/Agarose_Gel_Electrophoresis_and_DNA_Gel_Purification.pdf) |
| 18/08  * Assemblies [BII](https://static.igem.org/mediawiki/2014/4/48/BII.pdf) and   [BIII](https://static.igem.org/mediawiki/2014/e/ea/BIII.pdf):  + [Ligation](https://static.igem.org/mediawiki/2014/7/7a/Cohesive-end_assembly_cloning.pdf)  * Prepare LB medium (liquid and solid) | 19/08  * Assemblies [BII](https://static.igem.org/mediawiki/2014/4/48/BII.pdf) and   [BIII](https://static.igem.org/mediawiki/2014/e/ea/BIII.pdf):  + [Transformation](https://static.igem.org/mediawiki/2014/a/a5/Transformation_in_Escherichia_coli_DH5.pdf) | 20/08  * RBS+qteE, RBS+lasR and Assemblies [BII](https://static.igem.org/mediawiki/2014/4/48/BII.pdf) and   [BIII](https://static.igem.org/mediawiki/2014/e/ea/BIII.pdf):  + Inoculum | 21/08  * RBS+qteE, RBS+lasR and Assemblies [BII](https://static.igem.org/mediawiki/2014/4/48/BII.pdf) and   [BIII](https://static.igem.org/mediawiki/2014/e/ea/BIII.pdf):  + Glycerol Stock + [Miniprep](https://static.igem.org/mediawiki/2014/6/6f/Purelink_quick_plasmid_qrc.pdf) + [Restriction analysis](https://static.igem.org/mediawiki/2014/4/42/Restriction_Anaysis_and_Digestion_in_large_scale.pdf) | 22/08  * Prepare LB medium (liquid and solid) * Assembly [BII](https://static.igem.org/mediawiki/2014/4/48/BII.pdf):  + Repeat [restriction analysis](https://static.igem.org/mediawiki/2014/4/42/Restriction_Anaysis_and_Digestion_in_large_scale.pdf) adding the enzyme NdeI. | 23/08 | 24/08 |
| 25/08  * Inoculum  + [BBa\_J04450](http://parts.igem.org/Part%3ABBa_J04450) + [BBa\_K143055](http://parts.igem.org/Part%3ABBa_K143055)  * [PCR](https://static.igem.org/mediawiki/2014/7/74/PCR.pdf) [BBa\_K143055](http://parts.igem.org/Part%3ABBa_K143055) with higher melting temperature. | 26/08  * Assemblies [AI](https://static.igem.org/mediawiki/2014/b/b7/AI.pdf),   [CII](https://static.igem.org/mediawiki/2014/a/a3/CII.pdf),   [KIII](https://static.igem.org/mediawiki/2014/7/78/KIII.pdf),   [KIV](https://static.igem.org/mediawiki/2014/9/95/KIV.pdf) and   [KVI](https://static.igem.org/mediawiki/2014/8/83/KVI.pdf)  + [Digestion EXSP](https://static.igem.org/mediawiki/2014/4/42/Restriction_Anaysis_and_Digestion_in_large_scale.pdf) | 27/08  * Assemblies [AI](https://static.igem.org/mediawiki/2014/b/b7/AI.pdf),   [CII](https://static.igem.org/mediawiki/2014/a/a3/CII.pdf),   [KIII](https://static.igem.org/mediawiki/2014/7/78/KIII.pdf),   [KIV](https://static.igem.org/mediawiki/2014/9/95/KIV.pdf) and   [KVI](https://static.igem.org/mediawiki/2014/8/83/KVI.pdf)  + [Gel Purification](https://static.igem.org/mediawiki/2014/a/af/Agarose_Gel_Electrophoresis_and_DNA_Gel_Purification.pdf) | 28/08 | 29/08 | 30/08 | 31/08 |

## September

| Monday | Tuesday | Wednesday | Thursday | Friday | Saturday | Sunday |
| --- | --- | --- | --- | --- | --- | --- |
| 01/09  * Assembly [CII](https://static.igem.org/mediawiki/2014/a/a3/CII.pdf)   + [Digestion](https://static.igem.org/mediawiki/2014/4/42/Restriction_Anaysis_and_Digestion_in_large_scale.pdf) of the part BII with EcoRV.* Assemblies [AI](https://static.igem.org/mediawiki/2014/b/b7/AI.pdf),     [CII](https://static.igem.org/mediawiki/2014/a/a3/CII.pdf),     [KIII](https://static.igem.org/mediawiki/2014/7/78/KIII.pdf),     [KIV](https://static.igem.org/mediawiki/2014/9/95/KIV.pdf) and     [KVI](https://static.igem.org/mediawiki/2014/8/83/KVI.pdf):   + [Ligation](https://static.igem.org/mediawiki/2014/7/7a/Cohesive-end_assembly_cloning.pdf) | 02/09  * Assemblies [AI](https://static.igem.org/mediawiki/2014/b/b7/AI.pdf),   [CII](https://static.igem.org/mediawiki/2014/a/a3/CII.pdf),   [KIII](https://static.igem.org/mediawiki/2014/7/78/KIII.pdf),   [KIV](https://static.igem.org/mediawiki/2014/9/95/KIV.pdf) and   [KVI](https://static.igem.org/mediawiki/2014/8/83/KVI.pdf):  + [Transformation](https://static.igem.org/mediawiki/2014/a/a5/Transformation_in_Escherichia_coli_DH5.pdf) | 03/09  * Assemblies [AI](https://static.igem.org/mediawiki/2014/b/b7/AI.pdf),   [CII](https://static.igem.org/mediawiki/2014/a/a3/CII.pdf),   [KIII](https://static.igem.org/mediawiki/2014/7/78/KIII.pdf),   [KIV](https://static.igem.org/mediawiki/2014/9/95/KIV.pdf) and   [KVI](https://static.igem.org/mediawiki/2014/8/83/KVI.pdf):  + Inoculum | 04/09  * Assemblies [AI](https://static.igem.org/mediawiki/2014/b/b7/AI.pdf),   [CII](https://static.igem.org/mediawiki/2014/a/a3/CII.pdf),   [KIII](https://static.igem.org/mediawiki/2014/7/78/KIII.pdf),   [KIV](https://static.igem.org/mediawiki/2014/9/95/KIV.pdf) and   [KVI](https://static.igem.org/mediawiki/2014/8/83/KVI.pdf):  + Glycerol Stock + [Miniprep](https://static.igem.org/mediawiki/2014/6/6f/Purelink_quick_plasmid_qrc.pdf) + [Restriction analysis](https://static.igem.org/mediawiki/2014/4/42/Restriction_Anaysis_and_Digestion_in_large_scale.pdf) | 05/09 | 06/09 | 07/09 |
| 08/09  * [QuickChange PCR](https://static.igem.org/mediawiki/2014/6/63/QuickChange_PCR.pdf) to mutate the cleavage site of the [BBa\_K316037](http://parts.igem.org/Part%3ABBa_K316037) to a Cathepsin S recognition site * [Transformation](https://static.igem.org/mediawiki/2014/a/a5/Transformation_in_Escherichia_coli_DH5.pdf) of the QuickCHange PCR product | 09/09 | 10/09  * The QuickChange protocol didn't work well. So try again with more attention! * [QuickChange PCR](https://static.igem.org/mediawiki/2014/6/63/QuickChange_PCR.pdf) to mutate the cleavage site of the [BBa\_K316037](http://parts.igem.org/Part%3ABBa_K316037) to a Cathepsin S recognition site * [Transformation](https://static.igem.org/mediawiki/2014/a/a5/Transformation_in_Escherichia_coli_DH5.pdf) of the QuickCHange PCR product * Assemblies [DI](https://static.igem.org/mediawiki/2014/7/72/DI.pdf),   [KVIII](https://static.igem.org/mediawiki/2014/c/ce/KVIII.pdf),   [KX](https://static.igem.org/mediawiki/2014/9/95/KX.pdf) and   [KXI](https://static.igem.org/mediawiki/2014/9/9c/KXI.pdf):  + [Digestion EXSP](https://static.igem.org/mediawiki/2014/4/42/Restriction_Anaysis_and_Digestion_in_large_scale.pdf) | 11/09  * Prepare inoculum of the mutated [BBa\_K316037](http://parts.igem.org/Part%3ABBa_K316037) * Assemblies [DI](https://static.igem.org/mediawiki/2014/7/72/DI.pdf),   [KVIII](https://static.igem.org/mediawiki/2014/c/ce/KVIII.pdf),   [KX](https://static.igem.org/mediawiki/2014/9/95/KX.pdf) and   [KXI](https://static.igem.org/mediawiki/2014/9/9c/KXI.pdf):  + [Gel Purification](https://static.igem.org/mediawiki/2014/a/af/Agarose_Gel_Electrophoresis_and_DNA_Gel_Purification.pdf) + [Ligation](https://static.igem.org/mediawiki/2014/7/7a/Cohesive-end_assembly_cloning.pdf) | 12/09  * Assemblies [DI](https://static.igem.org/mediawiki/2014/7/72/DI.pdf),   [KVIII](https://static.igem.org/mediawiki/2014/c/ce/KVIII.pdf),   [KX](https://static.igem.org/mediawiki/2014/9/95/KX.pdf) and   [KXI](https://static.igem.org/mediawiki/2014/9/9c/KXI.pdf):  + [Transformation](https://static.igem.org/mediawiki/2014/a/a5/Transformation_in_Escherichia_coli_DH5.pdf)  * We had far to many difficulties trying to remove the RBS from the [BBa\_K143055](http://parts.igem.org/Part%3ABBa_K143055), so we decided to synthesize it using PCR. * [PCR for synthesis](https://static.igem.org/mediawiki/2014/3/35/PCR_for_synthesis.pdf) of the [BBa\_K143015](http://parts.igem.org/Part%3ABBa_K143015) ([BBa\_K143055](http://parts.igem.org/Part%3ABBa_K143055) without RBS) | 13/09  * Assembly [KX](https://static.igem.org/mediawiki/2014/9/95/KX.pdf)   + Inoculum* Assemblies [DI](https://static.igem.org/mediawiki/2014/7/72/DI.pdf),     [KVIII](https://static.igem.org/mediawiki/2014/c/ce/KVIII.pdf) and     [KXI](https://static.igem.org/mediawiki/2014/9/9c/KXI.pdf):   + Repeat [Ligation](https://static.igem.org/mediawiki/2014/7/7a/Cohesive-end_assembly_cloning.pdf)* Prepare LB medium (solid and liquid) | 14/09  * Assembly [KX](https://static.igem.org/mediawiki/2014/9/95/KX.pdf)   + Glycerol Stock   + [Miniprep](https://static.igem.org/mediawiki/2014/6/6f/Purelink_quick_plasmid_qrc.pdf)* Assemblies [DI](https://static.igem.org/mediawiki/2014/7/72/DI.pdf),     [KVIII](https://static.igem.org/mediawiki/2014/c/ce/KVIII.pdf) and     [KXI](https://static.igem.org/mediawiki/2014/9/9c/KXI.pdf):   + [Transformation](https://static.igem.org/mediawiki/2014/a/a5/Transformation_in_Escherichia_coli_DH5.pdf) |
| 15/09  * Assembly [KX](https://static.igem.org/mediawiki/2014/9/95/KX.pdf)   + [Restriction analysis](https://static.igem.org/mediawiki/2014/4/42/Restriction_Anaysis_and_Digestion_in_large_scale.pdf)* Assemblies [DI](https://static.igem.org/mediawiki/2014/7/72/DI.pdf),     [KVIII](https://static.igem.org/mediawiki/2014/c/ce/KVIII.pdf) and     [KXI](https://static.igem.org/mediawiki/2014/9/9c/KXI.pdf):   + Inoculum* [Gel Purification](https://static.igem.org/mediawiki/2014/a/af/Agarose_Gel_Electrophoresis_and_DNA_Gel_Purification.pdf) of the [BBa\_K143015](http://parts.igem.org/Part%3ABBa_K143015) | 16/09  * Prepare 40% glycerol solution * Assemblies [DI](https://static.igem.org/mediawiki/2014/7/72/DI.pdf),   [KVIII](https://static.igem.org/mediawiki/2014/c/ce/KVIII.pdf) and   [KXI](https://static.igem.org/mediawiki/2014/9/9c/KXI.pdf):  + Glycerol Stock + [Miniprep](https://static.igem.org/mediawiki/2014/6/6f/Purelink_quick_plasmid_qrc.pdf) + [Restriction analysis](https://static.igem.org/mediawiki/2014/4/42/Restriction_Anaysis_and_Digestion_in_large_scale.pdf) | 17/09  * [Ligation](https://static.igem.org/mediawiki/2014/0/03/PCR_Product_Clonig.pdf) of the synthesized [BBa\_K143015](http://parts.igem.org/Part%3ABBa_K143015) in TOPO vector followed by  [Transformation](https://static.igem.org/mediawiki/2014/a/a5/Transformation_in_Escherichia_coli_DH5.pdf)  * Measurement Interlab Study - Samples growth biobrick devices 1, 2 and 3 | 18/09  * Replate [BBa\_K143015](http://parts.igem.org/Part%3ABBa_K143015). We forgot the X-gal :(  * Measurement Interlab Study - Sample preparation and characterization by Flow Cytometry and Fluorometry | 19/09 | 20/09 | 21/09 |
| 22/09  * Inoculum of the [BBa\_K143015](http://parts.igem.org/Part%3ABBa_K143015) * Assembly [KXIV](https://static.igem.org/mediawiki/2014/6/67/KXIV.pdf):  + [Digestion EXSP](https://static.igem.org/mediawiki/2014/4/42/Restriction_Anaysis_and_Digestion_in_large_scale.pdf) | 23/09  * Assembly [KXIV](https://static.igem.org/mediawiki/2014/6/67/KXIV.pdf):  + [Digestion EXSP](https://static.igem.org/mediawiki/2014/4/42/Restriction_Anaysis_and_Digestion_in_large_scale.pdf)  * [Miniprep](https://static.igem.org/mediawiki/2014/6/6f/Purelink_quick_plasmid_qrc.pdf) and [Restriction Analysis](https://static.igem.org/mediawiki/2014/4/42/Restriction_Anaysis_and_Digestion_in_large_scale.pdf) of the [BBa\_K143015](http://parts.igem.org/Part%3ABBa_K143015) | 24/09  * Assembly [KXIV](https://static.igem.org/mediawiki/2014/6/67/KXIV.pdf):  + [Gel Purification](https://static.igem.org/mediawiki/2014/a/af/Agarose_Gel_Electrophoresis_and_DNA_Gel_Purification.pdf) + [Ligation](https://static.igem.org/mediawiki/2014/7/7a/Cohesive-end_assembly_cloning.pdf) + [Transformation](https://static.igem.org/mediawiki/2014/a/a5/Transformation_in_Escherichia_coli_DH5.pdf) | 25/09  * Assembly [KXIV](https://static.igem.org/mediawiki/2014/6/67/KXIV.pdf):  + Inoculum | 26/09  * Assembly [KXIV](https://static.igem.org/mediawiki/2014/6/67/KXIV.pdf):  + Glycerol Stock + [Miniprep](https://static.igem.org/mediawiki/2014/6/6f/Purelink_quick_plasmid_qrc.pdf) + [Restriction analysis](https://static.igem.org/mediawiki/2014/4/42/Restriction_Anaysis_and_Digestion_in_large_scale.pdf)  * Assemnlies [KII](https://static.igem.org/mediawiki/2014/b/b3/KII.pdf),   [KV](https://static.igem.org/mediawiki/2014/3/3d/KV.pdf) and   [AIV](https://static.igem.org/mediawiki/2014/e/eb/AIV.pdf):  + [Digestion EXSP](https://static.igem.org/mediawiki/2014/4/42/Restriction_Anaysis_and_Digestion_in_large_scale.pdf) | 27/09 | 28/09 |
| 29/09  * Assemnlies [KII](https://static.igem.org/mediawiki/2014/b/b3/KII.pdf),   [KV](https://static.igem.org/mediawiki/2014/3/3d/KV.pdf) and   [AIV](https://static.igem.org/mediawiki/2014/e/eb/AIV.pdf):  + [Gel Purification](https://static.igem.org/mediawiki/2014/a/af/Agarose_Gel_Electrophoresis_and_DNA_Gel_Purification.pdf) + [Ligation](https://static.igem.org/mediawiki/2014/7/7a/Cohesive-end_assembly_cloning.pdf) + [Transformation](https://static.igem.org/mediawiki/2014/a/a5/Transformation_in_Escherichia_coli_DH5.pdf) | 30/09  * Assemnlies [KII](https://static.igem.org/mediawiki/2014/b/b3/KII.pdf),   [KV](https://static.igem.org/mediawiki/2014/3/3d/KV.pdf) and   [AIV](https://static.igem.org/mediawiki/2014/e/eb/AIV.pdf):  + Inoculum |  |  |  |  |  |

## October

| Monday | Tuesday | Wednesday | Thursday | Friday | Saturday | Sunday |
| --- | --- | --- | --- | --- | --- | --- |
|  |  | 01/10  * Assemnlies [KII](https://static.igem.org/mediawiki/2014/b/b3/KII.pdf),   [KV](https://static.igem.org/mediawiki/2014/3/3d/KV.pdf) and   [AIV](https://static.igem.org/mediawiki/2014/e/eb/AIV.pdf):  + Glycerol Stock + [Miniprep](https://static.igem.org/mediawiki/2014/6/6f/Purelink_quick_plasmid_qrc.pdf) | 02/10 | 03/10 | 04/10 | 05/10 |
| 06/10  * Assemnlies [KII](https://static.igem.org/mediawiki/2014/b/b3/KII.pdf),   [KV](https://static.igem.org/mediawiki/2014/3/3d/KV.pdf) and   [AIV](https://static.igem.org/mediawiki/2014/e/eb/AIV.pdf):  + [Restriction analysis](https://static.igem.org/mediawiki/2014/4/42/Restriction_Anaysis_and_Digestion_in_large_scale.pdf)  * Assemblies [KIX](https://static.igem.org/mediawiki/2014/d/da/KIX.pdf),   [BIV](https://static.igem.org/mediawiki/2014/5/5b/BIV.pdf) and   KXVI  + [Digestion EXSP](https://static.igem.org/mediawiki/2014/4/42/Restriction_Anaysis_and_Digestion_in_large_scale.pdf) | 07/10  * Assemblies [KIX](https://static.igem.org/mediawiki/2014/d/da/KIX.pdf),   [BIV](https://static.igem.org/mediawiki/2014/5/5b/BIV.pdf) and   [KXVI](https://static.igem.org/mediawiki/2014/6/64/KXVI.pdf)  + [Gel Purification](https://static.igem.org/mediawiki/2014/a/af/Agarose_Gel_Electrophoresis_and_DNA_Gel_Purification.pdf) + [Ligation](https://static.igem.org/mediawiki/2014/7/7a/Cohesive-end_assembly_cloning.pdf) | 08/10  * Assemblies [KIX](https://static.igem.org/mediawiki/2014/d/da/KIX.pdf),   [BIV](https://static.igem.org/mediawiki/2014/5/5b/BIV.pdf) and   [KXVI](https://static.igem.org/mediawiki/2014/6/64/KXVI.pdf)  + [Transformation](https://static.igem.org/mediawiki/2014/a/a5/Transformation_in_Escherichia_coli_DH5.pdf) | 09/10- Assemblies [KIX](https://static.igem.org/mediawiki/2014/d/da/KIX.pdf),   [BIV](https://static.igem.org/mediawiki/2014/5/5b/BIV.pdf) and   [KXVI](https://static.igem.org/mediawiki/2014/6/64/KXVI.pdf)  * Inoculum | 10/10  * Assemblies [KIX](https://static.igem.org/mediawiki/2014/d/da/KIX.pdf),   [BIV](https://static.igem.org/mediawiki/2014/5/5b/BIV.pdf) and   [KXVI](https://static.igem.org/mediawiki/2014/6/64/KXVI.pdf)  + [Restriction analysis](https://static.igem.org/mediawiki/2014/4/42/Restriction_Anaysis_and_Digestion_in_large_scale.pdf) | 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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