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Team:Paris Saclay/Notebook/August/14
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[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=== |
''by Melanie'' | ''by Melanie'' | ||
| - | From the 20 ml of odor free culture (pJBEI Apra) done yesterday (''by Laetitia'') | + | From the 20 ml of odor free culture (pJBEI Apra - pPSI) done yesterday (''by Laetitia'') |
| - | + | Plasmid extraction without Nucleospin | |
| + | [https://2014.igem.org/Team:Paris_Saclay/Protocols/Extraction_of_the_Genomic_DNA_from_Bacteria_without_NucleoSpin%C2%AE_Tissue protocol] | ||
| + | + Stock (DH5a pJBEI + apra) | ||
| + | === Transformation of DH5α === | ||
| + | ''by Sean & Eugene '' | ||
| + | |||
| + | (Cacl2 protocol) | ||
| - | |||
| - | |||
We prepare a LB agar environment with a concentration of: | We prepare a LB agar environment with a concentration of: | ||
* 0.5 mM of IPTG | * 0.5 mM of IPTG | ||
| Line 29: | Line 33: | ||
and we spread our preparation in a petri dish and incubate overnight at 37°C. | and we spread our preparation in a petri dish and incubate overnight at 37°C. | ||
| - | + | ==Human Practices== | |
| - | + | ||
| - | + | ||
| - | + | ||
| - | ==Human | + | |
===Art & Design=== | ===Art & Design=== | ||
''by Terry'' | ''by Terry'' | ||
| Line 40: | Line 40: | ||
Preparation of M63 medium, concentration 5X, one liter basis : | Preparation of M63 medium, concentration 5X, one liter basis : | ||
| - | *10g ( | + | *10g (NH<sub>4</sub>)<sub>2</sub> SO<sub>4</sub> |
| - | *68g | + | *68g KH<sub>2</sub> PO<sub>4</sub> |
| - | *2.5mg | + | *2.5mg FeSO<sub>4</sub> 7H<sub>2</sub>O |
*Adjust to pH 7 with KOH | *Adjust to pH 7 with KOH | ||
The concentrated medium is diluted to 1X with sterile water and the following solution ( one liter basis ) : | The concentrated medium is diluted to 1X with sterile water and the following solution ( one liter basis ) : | ||
| - | *1 ml 1M | + | *1 ml 1M MgSO<sub>4</sub> 7H<sub>2</sub>O |
| - | *10 ml 20% carbon source ( sugar or glycerol ) | + | *10 ml 20% carbon source (sugar or glycerol) |
*0.1 ml 0.5% vitamin B1 | *0.1 ml 0.5% vitamin B1 | ||
| - | 100 ml of M63 has been prepared for | + | 100 ml of M63 has been prepared for future manipulations. |
| + | |||
| + | ==Photo of the Day== | ||
| + | [[File:Paris Saclay 14_august.jpg|600px|center]] | ||
| + | '''Members present:''' | ||
| + | * Instructors and advisors: Alice, Solenne and Sylvie. | ||
| + | * Students: Eugène, Fabio, Hoang Vu, Mélanie, Romain, Sean and Terry. | ||
{{Team:Paris_Saclay/notebook_footer}} | {{Team:Paris_Saclay/notebook_footer}} | ||
Latest revision as of 14:52, 14 October 2014
Contents |
Thursday 14th August
Lab Work
by Melanie
Preparation of DH5a CaCl2 Competent cell
Lemon Scent
by Melanie
From the 20 ml of odor free culture (pJBEI Apra - pPSI) done yesterday (by Laetitia)
Plasmid extraction without Nucleospin protocol
+ Stock (DH5a pJBEI + apra)
Transformation of DH5α
by Sean & Eugene
(Cacl2 protocol)
We prepare a LB agar environment with a concentration of:
- 0.5 mM of IPTG
- 0.008% of Xgal
- 1/2000 of Ampicilline
and we spread our preparation in a petri dish and incubate overnight at 37°C.
Human Practices
Art & Design
by Terry
We found an interesting colorless medium named M63.
Preparation of M63 medium, concentration 5X, one liter basis :
- 10g (NH4)2 SO4
- 68g KH2 PO4
- 2.5mg FeSO4 7H2O
- Adjust to pH 7 with KOH
The concentrated medium is diluted to 1X with sterile water and the following solution ( one liter basis ) :
- 1 ml 1M MgSO4 7H2O
- 10 ml 20% carbon source (sugar or glycerol)
- 0.1 ml 0.5% vitamin B1
100 ml of M63 has been prepared for future manipulations.
Photo of the Day
Members present:
- Instructors and advisors: Alice, Solenne and Sylvie.
- Students: Eugène, Fabio, Hoang Vu, Mélanie, Romain, Sean and Terry.
