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Team:Paris Saclay/Notebook/August/21
From 2014.igem.org
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=Thursday 21st August= | =Thursday 21st August= | ||
==Lab work== | ==Lab work== | ||
| - | === | + | ===Construction of the fusion protein (color)=== |
| - | ====Transformation of DH5α with chromoprotein==== | + | ====Transformation of DH5α with chromoprotein (in pGMETeasy)==== |
''by Sean'' | ''by Sean'' | ||
| + | Petri dishes were IPTG Xgal Amp | ||
[https://2014.igem.org/Team:Paris_Saclay/Protocols/Transformation_of_competent_e.coli_by_CaCl2 protocol] | [https://2014.igem.org/Team:Paris_Saclay/Protocols/Transformation_of_competent_e.coli_by_CaCl2 protocol] | ||
| - | === | + | ===Lemon Scent=== |
====Transformation of DH5α with CAD==== | ====Transformation of DH5α with CAD==== | ||
''by Sean'' | ''by Sean'' | ||
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==== Electro elution of pPS1 digested by Pac1 ==== | ==== Electro elution of pPS1 digested by Pac1 ==== | ||
| + | |||
| + | [[File:Paris_Saclay_140821.jpg|left]] | ||
| + | |||
''by Huang vu and laetitia'' | ''by Huang vu and laetitia'' | ||
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[https://2014.igem.org/Team:Paris_Saclay/Protocols/Electro_extraction_of_DNA_from_agarose_gel protocol of electro elution] | [https://2014.igem.org/Team:Paris_Saclay/Protocols/Electro_extraction_of_DNA_from_agarose_gel protocol of electro elution] | ||
| + | |||
| + | ''Gel electrophoresis'' | ||
| + | |||
| + | # ladder SM0403 5µl | ||
| + | # BBa_K762100 10µl | ||
| + | # BBa_K517003 10µl | ||
| + | # p cola 10µl | ||
| + | # pPSI 10µl | ||
====Extraction of pPSI==== | ====Extraction of pPSI==== | ||
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We have made an electrophoresis of purified CAD and pMCS5. | We have made an electrophoresis of purified CAD and pMCS5. | ||
| - | + | ||
| + | '''Dephosphorylation of pMCS5''' | ||
| + | |||
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30 min incubation in 37°c | 30 min incubation in 37°c | ||
| - | + | 15min of inactivation 65°c | |
| - | + | '''Ligation''' | |
| + | |||
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[https://2014.igem.org/Team:Paris_Saclay/Protocols/Electro_extraction_of_DNA_from_agarose_gel protocol] | [https://2014.igem.org/Team:Paris_Saclay/Protocols/Electro_extraction_of_DNA_from_agarose_gel protocol] | ||
| + | |||
| + | ==Photo of the Day== | ||
| + | [[File:Paris Saclay 21_august.jpg|600px|center]] | ||
| + | |||
| + | '''Members present:''' | ||
| + | * Instructors and advisors: Alice, Solenne and Sylvie. | ||
| + | * Students: Eugène, Hoang Vu, Laëtitia, Mélanie, Romain, Sean and Terry. | ||
{{Team:Paris_Saclay/notebook_footer}} | {{Team:Paris_Saclay/notebook_footer}} | ||
Latest revision as of 15:07, 14 October 2014
Contents |
Thursday 21st August
Lab work
Construction of the fusion protein (color)
Transformation of DH5α with chromoprotein (in pGMETeasy)
by Sean
Petri dishes were IPTG Xgal Amp protocol
Lemon Scent
Transformation of DH5α with CAD
by Sean
Electro elution of pPS1 digested by Pac1
by Huang vu and laetitia
We extracted the DNA from the agarose band saves yesterday
Gel electrophoresis
- ladder SM0403 5µl
- BBa_K762100 10µl
- BBa_K517003 10µl
- p cola 10µl
- pPSI 10µl
Extraction of pPSI
By Mélanie
From preculture of bacteria with pPSI we extract the plasmid
Extraction of pGMETeasy with LS GS and PS
By Mélanie
Following the cloning made by Laeticia in pGMETeasy, i extract the plasmid (protocol)
and I do some stock of each stain
Ligation of CAD in pMCS5
by Eugene
We have made an electrophoresis of purified CAD and pMCS5.
Dephosphorylation of pMCS5
| component | volume |
|---|---|
| H2O | 3 μl |
| pMCS5p | 5 μl |
| 10x buffer | 10 μl |
| Fast AP | 1 μl |
30 min incubation in 37°c
15min of inactivation 65°c
Ligation
| component | volume |
|---|---|
| CADp | 10 μl |
| pMCS5p | 2.5 μl |
| 10x buffer | 1.5 μl |
| Ligase | 1 μl |
night at 16°c
Photo of the Day
Members present:
- Instructors and advisors: Alice, Solenne and Sylvie.
- Students: Eugène, Hoang Vu, Laëtitia, Mélanie, Romain, Sean and Terry.
