Team:Paris Saclay/Notebook/June/30

From 2014.igem.org

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{{Team:Paris_Saclay/notebook_header}}
=Monday 30th June=
=Monday 30th June=
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==Lab work==
==Lab work==
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=== The odor-free ''E. coli'' chassis===
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====1 - Culture or the supercompetents:====
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====Culture of the supercompetents====
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''by Mathias''
''by Mathias''
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* MG1655Z1
* MG1655Z1
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1mL of an overnight culture in 100mL of LB.
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1ml of an overnight culture in 100ml of LB.
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====2 - Réhydration of selected biobricks:====
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====Rehydration of selected biobricks====
''by Juliette''
''by Juliette''
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* BBQ_I9026
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* BBa_I9026 (rhII translational unit)
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* BBQ_K592025
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* BBa_K592025 (BB0034-amilCP)
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* BBQ_K786002
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* BBa_K786002 (sensory rhodopsin II & Tar sensitive to blue-green light)
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* BBQ_K786003
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* BBa_K786003 (sensory rhodopsin I & Tar sensitive to orange light)
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* BBQ_K778002
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* BBa_K778002
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* BBQ_K592011 {2 times}
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* BBa_K592011 (cjBlue green chromoprotein)(2 times)
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* BBQ_K592009
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* BBa_K592009 (amilCP blue chromoprotein)
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* BBQ_K1033910 {idem}
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* BBa_K103391 (Yellow chromoprotein and Lime yellow-green chromoprotein)(2 times)
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* BBQ_J45014
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* BBa_J45014 (lemon scent, alcohol acetyltransferase I)
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* BBQ_K517003
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* BBa_K517003 (lemon scent, β-pinene)
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====3 - Transformations :====
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====Transformations====
''by Maher & Mathias''
''by Maher & Mathias''
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We use the DH5α provided by our advisors for the aforementioned Biobricks.
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We use the ''E. coli'' DH5α supercompetent cells provided by our advisors for the aforementioned Biobricks.
Spread on petri dishes with the concentration 10<sup>-1</sup> and 10<sup>-2</sup>.
Spread on petri dishes with the concentration 10<sup>-1</sup> and 10<sup>-2</sup>.
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====4 - Preparations :====
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====Preparations====
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''by Marie and Mathieu''
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* Aliquots of antibiotics :
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* Aliquots of antibiotics
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''by Marie & Mathieu''
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Put 0.203g of Chloramphenicol in 10ml of ethanol. Aliquots of 1ml.
Put 0.203g of Chloramphenicol in 10ml of ethanol. Aliquots of 1ml.
(Do the same for Kanamycin and Ampicillin)
(Do the same for Kanamycin and Ampicillin)
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* Petri dishes
 
''by Romain''
''by Romain''
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Add 250µl of antibiotics in 250ml of LB. Pour in petri dishes.
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* Petri dishes (with antiobiotics)
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* Range of gelose with different concentration of agar
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Add 250µl of antibiotics in 250ml of LAG. Pour in Petri dishes.
''by Arnaud''
''by Arnaud''
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 +
* Range of gelose with different concentration of agar
Add in 6 bottles of 500ml 2g of LB (lysogeny broth), and in each bottle add successively 1500mg (15mg/ml), 2000mg (20mg/ml), 2500mg (25mg/ml), 3000mg (30mg/ml), 3500mg (35mg/ml) and 4000mg (40mg/ml) of agar. And just before autoclaving complete each bottle with 100ml of reverse osmosis-purified water.
Add in 6 bottles of 500ml 2g of LB (lysogeny broth), and in each bottle add successively 1500mg (15mg/ml), 2000mg (20mg/ml), 2500mg (25mg/ml), 3000mg (30mg/ml), 3500mg (35mg/ml) and 4000mg (40mg/ml) of agar. And just before autoclaving complete each bottle with 100ml of reverse osmosis-purified water.
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==Photo of the Day==
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[[File:Paris Saclay 30_june.jpg|400px|center]]
'''Members present:'''
'''Members present:'''
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* Instructors : Solenne.
* Instructors : Solenne.
* Students : Arnaud, Juliette, Maher, Marie, Mathias, Mathieu, Pierre, Romain.
* Students : Arnaud, Juliette, Maher, Marie, Mathias, Mathieu, Pierre, Romain.
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{{Team:Paris_Saclay/notebook_footer}}
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[https://2014.igem.org/Team:Paris_Saclay/Notebook Back to the calendar]
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Latest revision as of 15:46, 14 October 2014

Contents

Monday 30th June

Lab work

The odor-free E. coli chassis

Culture of the supercompetents

by Mathias

  • MG1655
  • MG1655Z1

1ml of an overnight culture in 100ml of LB.

Rehydration of selected biobricks

by Juliette

  • BBa_I9026 (rhII translational unit)
  • BBa_K592025 (BB0034-amilCP)
  • BBa_K786002 (sensory rhodopsin II & Tar sensitive to blue-green light)
  • BBa_K786003 (sensory rhodopsin I & Tar sensitive to orange light)
  • BBa_K778002
  • BBa_K592011 (cjBlue green chromoprotein)(2 times)
  • BBa_K592009 (amilCP blue chromoprotein)
  • BBa_K103391 (Yellow chromoprotein and Lime yellow-green chromoprotein)(2 times)
  • BBa_J45014 (lemon scent, alcohol acetyltransferase I)
  • BBa_K517003 (lemon scent, β-pinene)

Transformations

by Maher & Mathias

We use the E. coli DH5α supercompetent cells provided by our advisors for the aforementioned Biobricks. Spread on petri dishes with the concentration 10-1 and 10-2.

Preparations

by Marie and Mathieu

  • Aliquots of antibiotics

Put 0.203g of Chloramphenicol in 10ml of ethanol. Aliquots of 1ml. (Do the same for Kanamycin and Ampicillin)

by Romain

  • Petri dishes (with antiobiotics)

Add 250µl of antibiotics in 250ml of LAG. Pour in Petri dishes.

by Arnaud

  • Range of gelose with different concentration of agar

Add in 6 bottles of 500ml 2g of LB (lysogeny broth), and in each bottle add successively 1500mg (15mg/ml), 2000mg (20mg/ml), 2500mg (25mg/ml), 3000mg (30mg/ml), 3500mg (35mg/ml) and 4000mg (40mg/ml) of agar. And just before autoclaving complete each bottle with 100ml of reverse osmosis-purified water.

Photo of the Day

Paris Saclay 30 june.jpg

Members present:

  • Instructors : Solenne.
  • Students : Arnaud, Juliette, Maher, Marie, Mathias, Mathieu, Pierre, Romain.

Back to the calendar