Difference between revisions of "Team:Paris Saclay/Notebook/August/17"

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===
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====Culture of BL21====
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''By Charlène''
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3 clones from each transformation condition were grown in 500µL of LB, streptomycin (50µg/mL) and chloramphenicol (30µg/mL).
 +
Then each solution was split into 3x150µL and 350µL of mix of LB + streptomycin (50µg/mL) + chloramphenicol (30µg/mL) were added to each culture.
 +
Salicylate (main solution at 10mM) was added to obtain the following concentrations :
 +
 
 +
{| class="wikitable"
 +
|-
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!Plasmid(s)
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|colspan="9"|K1372001
 +
 
 +
|colspan="9"|K1372001 + pcl_TAA
 +
|colspan="9"|K1372001 + pcl_TAG
 +
|colspan="9"|K1372001 + pcl_Tq
 +
|-
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!Clone
 +
|1
 +
|2
 +
|3
 +
|1
 +
|2
 +
|3
 +
|1
 +
|2
 +
|3
 +
|1
 +
|2
 +
|3
 +
|1
 +
|2
 +
|3
 +
|1
 +
|2
 +
|3
 +
|1
 +
|2
 +
|3
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|1
 +
|2
 +
|3
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|1
 +
|2
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|3
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|1
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|2
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|3
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|1
 +
|2
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|3
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|1
 +
|2
 +
|3
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|-
 +
!Salicylate concentration
 +
|colspan="3"|0
 +
|colspan="3"|30µM
 +
|colspan="3"|1mM
 +
|colspan="3"|0
 +
|colspan="3"|30µM
 +
|colspan="3"|1mM
 +
|colspan="3"|0
 +
|colspan="3"|30µM
 +
|colspan="3"|1mM
 +
|colspan="3"|0
 +
|colspan="3"|30µM
 +
|colspan="3"|1mM
 +
|}
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 +
{{Team:Paris_Saclay/notebook_footer}}
  
 
{{Team:Paris_Saclay/notebook_footer}}
 
{{Team:Paris_Saclay/notebook_footer}}

Revision as of 15:17, 17 August 2016

Wednesday 17th August

Lab work

Visualization

Extraction of pPS16003, pPS16004 and GFP1-9_PSB1C3 (clones 2 and 3)

By Charlène

Clones 2 and 3 of GFP1-9_PSB1C3, and pPS16003, pPS16004 were extracted with the Plasmid MiniPrep kit.

The extracts were put to migrated on a 0.8%agarose gel with BET.

Purification of gBlocks 2, 3 and 4

By Terrence

The purification was carried out following the usual protocol.


Cloning with pJET the gBlock 1.2, FRB, FKBP and SgNM

By Léa and Naiane

The cloning was carried out using a new protocol which uses pJET as cloning vector.

A heat shock transformation was made on the cloning samples using the following protocol


Culture of BL21

By Charlène

3 clones from each transformation condition were grown in 500µL of LB, streptomycin (50µg/mL) and chloramphenicol (30µg/mL). Then each solution was split into 3x150µL and 350µL of mix of LB + streptomycin (50µg/mL) + chloramphenicol (30µg/mL) were added to each culture. Salicylate (main solution at 10mM) was added to obtain the following concentrations :

Plasmid(s) K1372001 K1372001 + pcl_TAA K1372001 + pcl_TAG K1372001 + pcl_Tq
Clone 1 2 3 1 2 3 1 2 3 1 2 3 1 2 3 1 2 3 1 2 3 1 2 3 1 2 3 1 2 3 1 2 3 1 2 3
Salicylate concentration 0 30µM 1mM 0 30µM 1mM 0 30µM 1mM 0 30µM 1mM





Université Paris Sud
91405 Orsay, France
Copyright © 2016 iGEM Paris Saclay. All rights reserved All inspirations encouraged.
*This wiki was conceived during long nights of work and a great coffee experience in the beautiful Orsay, France.





Université Paris Sud
91405 Orsay, France
Copyright © 2016 iGEM Paris Saclay. All rights reserved All inspirations encouraged.
*This wiki was conceived during long nights of work and a great coffee experience in the beautiful Orsay, France.