Team:Paris Saclay/Notebook/August/14

From 2014.igem.org

(Difference between revisions)
(D-Lemon Scent)
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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]
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===D-Lemon Scent===
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===Lemon Scent===
''by Melanie''
''by Melanie''
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From the 20 ml of odor free culture (pJBEI Apra) done yesterday (''by Laetitia'')
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From the 20 ml of odor free culture (pJBEI Apra - pPSI) done yesterday (''by Laetitia'')
Plasmid extraction without Nucleospin
Plasmid extraction without Nucleospin
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[https://2014.igem.org/Team:Paris_Saclay/Protocols/Extraction_of_the_Genomic_DNA_from_Bacteria_without_NucleoSpin%C2%AE_Tissue protocol]
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=== Transformation of DH5 alpha ===
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+ Stock (DH5a pJBEI + apra)
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=== Transformation of DH5α ===
''by Sean & Eugene ''
''by Sean & Eugene ''
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 +
(Cacl2 protocol)
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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  
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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.
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==Human Practices==
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[https://2014.igem.org/Team:Paris_Saclay/Protocols/Extraction_of_the_Genomic_DNA_from_Bacteria_without_NucleoSpin%C2%AE_Tissue Protocol]
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==Human Pratices==
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===Art & Design===
===Art & Design===
''by Terry''
''by Terry''
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Preparation of M63 medium, concentration 5X, one liter basis :
Preparation of M63 medium, concentration 5X, one liter basis :
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*10g (NH4)2 SO4
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*10g (NH<sub>4</sub>)<sub>2</sub> SO<sub>4</sub>
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*68g KH2 PO4
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*68g KH<sub>2</sub> PO<sub>4</sub>
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*2.5mg FeSO4 7H2O
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*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 MgSO4 7H2O
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*1 ml 1M MgSO<sub>4</sub> 7H<sub>2</sub>O
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*10 ml 20% carbon source ( sugar or glycerol )
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*10 ml 20% carbon source (sugar or glycerol)
*0.1 ml 0.5% vitamin B1
*0.1 ml 0.5% vitamin B1
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100 ml of M63 has been prepared for futur manipulations.
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100 ml of M63 has been prepared for future manipulations.
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==Photo of the Day==
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[[File:Paris Saclay 14_august.jpg|600px|center]]
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'''Members present:'''
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* Instructors and advisors: Alice, Solenne and Sylvie.
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* 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

Protocol

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

Paris Saclay 14 august.jpg

Members present:

  • Instructors and advisors: Alice, Solenne and Sylvie.
  • Students: Eugène, Fabio, Hoang Vu, Mélanie, Romain, Sean and Terry.

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