Team:Evry/Notebook/w2

From 2012.igem.org

(Difference between revisions)
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<li>Xenopus embryos present 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.  
<li>Xenopus embryos present 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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<li>On 96 plates : <br>
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<li>On <bold>96 plates</bold>, each plate contains 1 embryo:
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'''Plate 1''' <br>
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each plate contain : 1 embryo <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
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<br>
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'''Plate 2'''
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normal embryos only
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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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<ul>
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<li>Plate 1:
 +
<ul>
 +
<li>Line A & B = on embryos without protective membrane : 500 uL mRFP plasmid on DH5a deposite + 1mL LB + 1mL MMR medium <br>
 +
<li>Line C : 1 2 3 : normal embryos : 500 uL mRFP plasmid on DH5a deposite + 1mL (LB + MMR medium) <br>
 +
<li>Line E & F = embryos without protective membrane : 500 uL AUX plasmid integrated on DH5a deposit + 1mL LB + 1mL MMR medium <br>
 +
<li>Line G & M1-6 = normal embryos : 500 uL AUX plasmid integrated on DH5a deposit + 1mL LB + 1mL MMR medium <br>
 +
<li>Line M7-9 = normal embryos : controls
 +
</ul>
 +
<li> Plate 2 (normal embryos only):
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<ul>
 +
<li>Line A & B
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<li>Line C : LB/MMR medium
 +
<li>Line D : RFP 0.1uL
 +
<li>Line E : AUX 0.1uL
 +
<li>Line G & H : LB/MMR medium only (no embryos)
 +
</ul>
<img src="https://static.igem.org/mediawiki/2012/8/8d/SNAP-175407-0003.jpg" alt="result example" />
<img src="https://static.igem.org/mediawiki/2012/8/8d/SNAP-175407-0003.jpg" alt="result example" />
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</ul>
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<li>On <bold>16 Plates</bold> each plate contains 3 tadpoles:
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<ul>
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<li>On '''16 Plates'''
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<li>Line A : 0.5uL RFP bacteria
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<br><br>
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<li>Line B1 : 0.3uL RFP bacteria
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Tadpoles experimentations : <br> ''each plate contains 3 tadpoles''
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</ul>
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<br>
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Line A : 0.5uL RFP bacteria <br>
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Line B1 : 0.3uL RFP bacteria
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<br>
<br>
</ol>
</ol>

Revision as of 19:49, 2 August 2012

Weeks

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16

Week 2: 18th June - 24th June

Monday, 18th June

  • Inoculation
  • Auxin and salkowski's test purchased

Tuesday, 19th June

Miniprep

  • J23100 RFP: 101.9 ng/uL
  • K515100 Auxin: 319.7 ng/uL

Misc

Liquid culture of aux and RFP

Wednesday, 20th June

Previous work :

Liquid culture of DH5a with mRFP and auxin plasmid

Main goals:

  1. Auxin toxicity test
  2. Bacterial integration assessment into the xenopus embryos

Experiments

  1. Xenopus embryos present 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.
  2. On 96 plates, each plate contains 1 embryo:
    • Plate 1:
      • Line A & B = on embryos without protective membrane : 500 uL mRFP plasmid on DH5a deposite + 1mL LB + 1mL MMR medium
      • Line C : 1 2 3 : normal embryos : 500 uL mRFP plasmid on DH5a deposite + 1mL (LB + MMR medium)
      • Line E & F = embryos without protective membrane : 500 uL AUX plasmid integrated on DH5a deposit + 1mL LB + 1mL MMR medium
      • Line G & M1-6 = normal embryos : 500 uL AUX plasmid integrated on DH5a deposit + 1mL LB + 1mL MMR medium
      • Line M7-9 = normal embryos : controls
    • Plate 2 (normal embryos only):
      • Line A & B
      • Line C : LB/MMR medium
      • Line D : RFP 0.1uL
      • Line E : AUX 0.1uL
      • Line G & H : LB/MMR medium only (no embryos)
      result example
  3. On 16 Plates each plate contains 3 tadpoles:
    • Line A : 0.5uL RFP bacteria
    • Line B1 : 0.3uL RFP bacteria

We put this plate on UV light 15min after the deposit: result example

Thursday, 21st June

Friday, 22nd June

Retrieved from "http://2012.igem.org/Team:Evry/Notebook/w2"