iGEM 2016 — Team Saint_Rose_School_A

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# Team:Saint Rose School A/Notebook

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* [Home](https://2016.igem.org/Team%3ASaint_Rose_School_A)
* [Project](https://2016.igem.org/Team%3ASaint_Rose_School_A/Description)
* [Notebook](https://2016.igem.org/Team%3ASaint_Rose_School_A/Notebook)
* [Results](https://2016.igem.org/Team%3ASaint_Rose_School_A/Results)
* [Safety](https://2016.igem.org/Team%3ASaint_Rose_School_A/Safety)
* [Human Practices](https://2016.igem.org/Team%3ASaint_Rose_School_A/Human_Practices)
* [Collaborations](https://2016.igem.org/Team%3ASaint_Rose_School_A/Collaborations)
* [Attributions](https://2016.igem.org/Team%3ASaint_Rose_School_A/Attributions)
* [Team](https://2016.igem.org/Team%3ASaint_Rose_School_A/Team)

# Notebook

- June-October
- **June**
- **Week 01:**
- **Friday 24:** ° Search BioBricks parts to use.
- **Saturday 25:** ° Search for available parts.

- **July**
- **Week 01:**
- **Friday 01:** ° Leave E.coli bacteria culture to generate chemocompetents (for processing). LuxR ° Cultivate bacteria with genes.
- **Saturday 02:** ° Removing purified plasmid LuxR- LuxI.

- **Week 02:**
- **Wednesday 06:**
   ° Learn spectrophotometry, concentration, purity and DNA restriction analysis.
  ° Check bacterial growth.
- **Friday 08:**
   Purified plasmid extraction and LuxR to know why there was an error with gen Luxi
   ° Electrophoresis with LuxI LuxR genes and without result

- **Week 03:**
- **Wednesday 13:**
   ° the cultivation LuxR was reviewed.
  ° We made again the purified plasmid with the gene extraction
   LuxR, since no crop LuxI gene.
  ° Extract, transform and cultivate LuxR, LuxI genes and Plux.
   \* Prior to transformation
- **Friday 15:**
   ° Review LuxI growth genes, LuxR, Plux
    They grew completely genes
   ° 09/07 LuxR growth gene was verified
   This gene was contaminated, it was not followed with procedure (red color)
    ° the extraction with purified plasmid genes was performed LuxI, LuxR and Plux.
   ° Then took electrophoresis to Colonial De Pirque School.
   We could see only the Plux, with the other two genes
   There was no result.
    The Colonial De Pirque School had result  LuxI LuxR and not with Plux.
   Plasmids were shared to further progress.

- **Week 04:**
- **Friday 22:**
   ° Cortes with Olux plasmids, LuxR, LuxI
   Shared school and worked with CCP
   [EcoR1, PstI] - buffer h
   ° After cutting waited 1hr
   ° After electrophoresis was more the previous week (plasmid)
   \* Camera girl: much heated and do not see anything.
   \* Large Camera: There was an error and failed to see.

- **Saturday 23:**
  electrophoresis ° was repeated with the same plasmids of the previous day
   -Plux 22/07
  -LuxI 22/07
   -LuxR 22/07
   -LuxR 15/07
   -LuxI 15/07
   -Plux 15/07 (CCP)
    \* Large Camera: -Ladder 1000 bp
     \* Camera girl:
   -first 1000 bp ladder
    -second 100 bp ladder
   Not able to see any of the two chambers
  ° Electrophoresis was done again with no digested plasmids.
   Plux: 15/07
   LuxR: 15/07 (CCP)
   LuxI: 15/07 (CCP)
   They were achieved see, as they were good
   ° They will digest / court to verify this all good

- **Friday 29:**
   ° cut with plasmid Plux 15/07, and also repeat LuxR
   CCP LuxI
   ° Javier made the cut with plasmids made. LuxR, LuxI and Plux
   ° Plux was electrophoreses
   He managed to see only one band

- **Saturday 30:**
   cut plasmids ° electrophoresis
   CCP LuxI
   (1.2 gel)
   1-. 29/07 Plux cut plasmid (see achieved)
   2-. 29/07 LuxR cut plasmid (see achieved)
   3-. 29/07 LuxI cut plasmid
   4-. 22/07 LuxR cut plasmid (see achieved)
   5-. 22/07 Plux cut plasmid (see achieved) 6. 22/07 LuxI cut plasmid
   ° electrophoresis extracted plasmids
  1-15 / 07 plasmid extracted LuxR
   2-15 / 07 plasmid extracted LuxI
   3-15 / 07 plasmid extracted Plux
   They were able to see, but they were very degraded, they were not clear

- **August**

- **Week 01**

- **Friday 5:**
  ° electrophoresis is performed again with the plasmids:
   -Plux
   -LuxR
    -LuxI
   They were able to see clearly, the second tends to have two bands
   ° It will grow with Luxi LuxR and bacteria Plux chloramphenicol
   ° is grown: - LuxR
   - LuxI
   - Plux1
   - Plux2

- **Saturday 06:**
  ° the Luxi, LuxR, Plux1 Plux2 be extracted and controlled

  **Week 2:**
  **-Wednesday 10:**
  ° digestion concentrates plasmids: -LuxR -Plux1
  -LuxI -Plux2
   ° Electrophoresis with plasmids concentrates

- **Friday 12:**
  He turned to make electrophoresis to chec

- **Saturday 13:**
  ° Cryogenics with Plux 1 Plux2, LuxI and LuxR plasmids.
   ° recultivate all Lux.

- **Week 03:**

  Saturday 20:
   chromoprotein ° Cultivate purple (AmilCP)

  **Week 04**

  **Wednesday 24:**

  ° Check cultivation
  It did not grow
  ° Make Calcium to transform competent
   ° He turned to transform and to cultivate

  **Friday 26:**
  ° plasmidial Removing the purple (AmilCP)

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