Team:Evry/Notebook/w2
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<h2>Monday, 18th June</h2> | <h2>Monday, 18th June</h2> | ||
+ | |||
+ | <ul> | ||
+ | <li>Inoculation | ||
+ | <li>Auxin and salkowski's test purchased </b> | ||
+ | </ul> | ||
<h2>Tuesday, 19th June</h2> | <h2>Tuesday, 19th June</h2> | ||
+ | <h3> Miniprep </h3> | ||
+ | <ul> | ||
+ | <li>J23100 RFP: 101.9 ng/uL <br> | ||
+ | <li>K515100 Auxin: 319.7 ng/uL <br> | ||
+ | </ul> | ||
+ | |||
+ | <h3>Misc</h3> | ||
+ | |||
+ | liquid culture of aux and RFP | ||
<h2>Wednesday, 20th June</h2> | <h2>Wednesday, 20th June</h2> | ||
+ | |||
+ | |||
+ | <h3>Previous work :</h3> | ||
+ | |||
+ | liquid culture of DH5a with mRFP and auxin plasmid | ||
+ | |||
+ | <h3>Main goals:</h3> | ||
+ | <ol> | ||
+ | <li>Auxin toxicity test | ||
+ | <li>Bacterial integration assessment into the xenopus embryos | ||
+ | </ol> | ||
+ | |||
+ | <h3>Experiments</h3> | ||
+ | |||
+ | <ol> | ||
+ | <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>On 96 plates : <br> | ||
+ | '''Plate 1''' <br> | ||
+ | each plate contain : 1 embryo <br> | ||
+ | |||
+ | Line A & B = on embryos without protective membrane : 500 uL mRFP plasmid on DH5a deposite + 1mL LB + 1mL MMR medium <br> | ||
+ | Line C : 1 2 3 : normal embryos : 500 uL mRFP plasmid on DH5a deposite + 1mL (LB + MMR medium) <br> | ||
+ | <br> | ||
+ | Line E & F = embryos without protective membrane : 500 uL AUX plasmid integrated on DH5a deposit + 1mL LB + 1mL MMR medium <br> | ||
+ | Line G & M1-6 = normal embryos : 500 uL AUX plasmid integrated on DH5a deposit + 1mL LB + 1mL MMR medium <br> | ||
+ | Line M7-9 = normal embryos : controls | ||
+ | <br> | ||
+ | |||
+ | |||
+ | '''Plate 2''' | ||
+ | normal embryos only | ||
+ | |||
+ | Line A & B <br> | ||
+ | Line C : LB/MMR medium <br> | ||
+ | Line D : RFP 0.1uL<br> | ||
+ | Line E : AUX 0.1uL<br> | ||
+ | Line G & H : LB/MMR medium only (no embryos)<br> | ||
+ | |||
+ | <img> src="https://static.igem.org/mediawiki/2012/8/8d/SNAP-175407-0003.jpg" alt="result example" /> | ||
+ | |||
+ | |||
+ | |||
+ | <li>On '''16 Plates''' | ||
+ | <br><br> | ||
+ | Tadpoles experimentations : <br> ''each plate contains 3 tadpoles'' | ||
+ | <br> | ||
+ | Line A : 0.5uL RFP bacteria <br> | ||
+ | Line B1 : 0.3uL RFP bacteria | ||
+ | <br> | ||
+ | </ol> | ||
+ | |||
+ | We put this plate on UV light 15min after the deposit: | ||
+ | |||
+ | <img src="https://static.igem.org/mediawiki/2012/a/a9/Wt-0001.jpg" alt="result example"/> | ||
<h2>Thursday, 21st June</h2> | <h2>Thursday, 21st June</h2> |
Revision as of 19:43, 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 RFPWednesday, 20th June
Previous work :
liquid culture of DH5a with mRFP and auxin plasmidMain goals:
- Auxin toxicity test
- Bacterial integration assessment into the xenopus embryos
Experiments
- 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.
- 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)
src="https://static.igem.org/mediawiki/2012/8/8d/SNAP-175407-0003.jpg" alt="result example" /> - On '''16 Plates'''
Tadpoles experimentations :
''each plate contains 3 tadpoles''
Line A : 0.5uL RFP bacteria
Line B1 : 0.3uL RFP bacteria