Team:Evry/Notebook/w2

From 2012.igem.org

(Difference between revisions)
Line 44: Line 44:
<h3>Misc</h3>
<h3>Misc</h3>
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liquid culture of aux and RFP
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Liquid culture of aux and RFP
<h2>Wednesday, 20th June</h2>
<h2>Wednesday, 20th June</h2>
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<h3>Previous work :</h3>
<h3>Previous work :</h3>
-
liquid culture of DH5a with mRFP and auxin plasmid
+
Liquid culture of DH5a with mRFP and auxin plasmid
<h3>Main goals:</h3>
<h3>Main goals:</h3>
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Line G & H : LB/MMR medium only (no embryos)<br>
Line G & H : LB/MMR medium only (no embryos)<br>
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<img> src="https://static.igem.org/mediawiki/2012/8/8d/SNAP-175407-0003.jpg" alt="result example" />
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<img src="https://static.igem.org/mediawiki/2012/8/8d/SNAP-175407-0003.jpg" alt="result example" />

Revision as of 19:44, 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 :
    '''Plate 1'''
    each plate contain : 1 embryo
    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'''

    Tadpoles experimentations :
    ''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