Team:Evry/Notebook/June/21

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(Created page with "Today's achievements: <i><b>Previous work :</b> liquid culture of DH5a with mRFP and auxin plasmid</i> Main goals : <br> °Auxin toxicity test <br> °Bacterial integration ass...")
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Today's achievements:
Today's achievements:
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<i><b>Previous work :</b> liquid culture of DH5a with mRFP and auxin plasmid</i>
 
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Main goals : <br>  
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Yesterday results <br>
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°Auxin toxicity test <br>
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°Bacterial integration assessment into the xenopus embryos
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1°/Xenopus embryos presents a protective membrane, we decided to realize our bacterial deposits experiments on xenopus with and without their membrane. Then, we could see if bacterias could enter into the embryos naturally.
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A = Alive ; D = Dead
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2°/ On 96 plates : <br>
 
'''Plate 1''' <br>
'''Plate 1''' <br>
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each plate contain : 1 embryo <br>
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All the embryos are dead except H7<br>
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Line A & B = on embryos without protective membrane : 500 uL mRFP plasmid on DH5a deposite + 1mL LB + 1mL MMR medium <br>
 
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Line C : 1 2 3 : normal embryos : 500 uL mRFP plasmid on DH5a deposite + 1mL (LB + MMR medium) <br>
 
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<br>
 
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Line E & F = embryos without protective membrane : 500 uL AUX plasmid integrated on DH5a deposit + 1mL LB + 1mL MMR medium <br>
 
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Line G & M1-6 = normal embryos : 500 uL AUX plasmid integrated on DH5a deposit + 1mL LB + 1mL MMR medium <br>
 
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Line M7-9 = normal embryos : controls
 
<br>
<br>
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'''Plate 2'''
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'''Plate 2'''<br><br>
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normal embryos only
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Line ABC : all dead
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Line D : 1D ; 2D ; 3A ; 4D ; 5A ; 6D ; 7-8-9A ; 10D ; 11-12A <br>
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Line E : 1A ; 2D ; 3D ; 4A ; 5D ; 6A ; 7-8-9-10D ; 11-12A
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Line A & B  <br>
 
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Line C : LB/MMR medium <br>
 
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Line D : RFP 0.1uL<br>
 
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Line E : AUX 0.1uL<br>
 
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Line G & H : LB/MMR medium only (no embryos)<br>
 
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https://static.igem.org/mediawiki/2012/8/8d/SNAP-175407-0003.jpg
 
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'''Plate 3'''<br>
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3°/ On '''16 Plates'''
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<br><br>
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Tadpoles experimentations : <br> ''each plate contains 3 tadpoles''
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<br>
<br>
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Line A : 0.5uL RFP bacteria <br>
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Line A : 1D ; 2-3-4A<br>
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Line B1 : 0.3uL RFP bacteria
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Line B1 : dead
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<br>
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We put this plate on UV light 15min after the deposit:
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https://static.igem.org/mediawiki/2012/a/a9/Wt-0001.jpg
 
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<i>Comments : <br>
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Plate 2 : 50% of dead when we spread bacterias on embryos. All dead if we drown the embryos of bacteria. <br>
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Plate 1 : All dead except 1 <br>
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Plate 3 : All the tadpoles are fluorescent except the control one.
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Revision as of 13:02, 10 July 2012

Today's achievements:


Yesterday results

A = Alive ; D = Dead

Plate 1
All the embryos are dead except H7



Plate 2

Line ABC : all dead Line D : 1D ; 2D ; 3A ; 4D ; 5A ; 6D ; 7-8-9A ; 10D ; 11-12A
Line E : 1A ; 2D ; 3D ; 4A ; 5D ; 6A ; 7-8-9-10D ; 11-12A


Plate 3

Line A : 1D ; 2-3-4A
Line B1 : dead


Comments :
Plate 2 : 50% of dead when we spread bacterias on embryos. All dead if we drown the embryos of bacteria.
Plate 1 : All dead except 1
Plate 3 : All the tadpoles are fluorescent except the control one.