Team:Saint Rose School A/Notebook
Notebook
° Learn spectrophotometry, concentration, purity and DNA restriction analysis.
° Check bacterial growth.
Purified plasmid extraction and LuxR to know why there was an error with gen Luxi
° Electrophoresis with LuxI LuxR genes and without result
° 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
° 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.
° 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.
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
° 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
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
° 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
° the Luxi, LuxR, Plux1 Plux2 be extracted and controlled
Week 2:
-Wednesday 10:
° digestion concentrates plasmids: -LuxR -Plux1
-LuxI -Plux2
° Electrophoresis with plasmids concentrates
He turned to make electrophoresis to chec
° Cryogenics with Plux 1 Plux2, LuxI and LuxR plasmids.
° recultivate all Lux.
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)
