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From 2012.igem.org

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Revision as of 12:50, 10 July 2012

Today's achievements:

Previous work : liquid culture of DH5a with mRFP and auxin plasmid

Main goals :
°Auxin toxicity test
°Bacterial integration assessment into the xenopus embryos

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.

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)

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: