Team:Paris Saclay/Notebook/August/5

Wednesday 5th August

Lab Work

Quantification

by Pauline and Coralie

Biobricks:

• BBa_R0051
• BBa_E0022 (15ng/mL)
• BBa_E0422 (20ng/mL)
• BBa_K1707000 (20ng/mL)
• BBa_K1707004 (15ng/mL)
• BBa_K1707011
• BBa_K1707012
• BBa_K1707013 (20ng/mL)
• BBa_K1707019 (15ng/mL)

Agarose gel, 1% Migration 90V

Ligation

by Pauline

Biobricks: BBa_K1707020 : BBa_K1707004 + BBa_K1707019

Mix:

• 6,5 µL BBa_K1707019
• 9,5 µL BBa_K1707004
• 2 µL Tampon 10x
• 1 µL Ligase
• 1 µL H2O

BBa Test : BBa_K1707004 + BBa_K1707013

Mix:

• 5 µL BBa_K1707013
• 7,5 µL BBa_K1707004
• 1,5 µL Tampon 10x
• 1 µL Ligase

BBa_K1707027 : BBa_K1707000 + BBa_E0022

Mix:

• 5 µL BBa_K1707000
• 9 µL BBa_E0022
• 1,5 µL Tampon 10x
• 1 µL Ligase
• 3,5 µL H2O

BBa_K1707028 : BBa_R0051 + BBa_E0422

Mix:

• 6,25 µL BBa_R0051
• 5,5 µL BBa_E0422
• 1,5 µL Tampon 10x
• 1 µL Ligase
• 0,75 µL H2O

Rehydratation

by Audrey

BBa_R0040

Transformation

by Audrey and Coralie

• BBa_R0040
• BBa Test
• BBa_K1707028
• BBa_K1707027
• BBa_K1707020

Digestion

by Pauline

XbaI + PstI:

• BBa_K1707012
• BBa_K1707011
• BBa_I13600

SpeI + Pst:

• BBa_R0051
• BBa_K1707008

EcoRI + SpeI:

• BBa_K1707004

XbaI + EcoRI:

• BBa_I13602

Mix:

• 10 µL Plasmide
• 1 µL Enzyme 1
• 1 µL Enzyme 2
• 2 µL Buffer FastDigest 10x
• 6 µL H2O

Soil experiment

by Coralie

Soil

Observation of plates (04/08/2015):

• Soil + H2O on new MCK: lots of colonies are visible
• Soil + LB on new MCK: lots of colonies are visible
• LB on new MCK: nothing
• MCK alone: nothing

We can suppose two different hypothesis:

• The soil is contaminated
• the MCK isn't OK for our experiment: now we add lactose in the preparation. We will try with sorbitol instead of lactose.

We try soil + H2O and soil +LB on:

• MCK + Sorbitol: Without antibiotics, with Spectinomycine, Tetramycine or Chloramphenicol
• MCK + Lactose: Without antibiotics, with Spectinomycine, Tetramycine or Chloramphenicol

Incubation ON, 37°C

Member present:

• Instructors: Claire
• Students: Coralie, Audrey and Pauline

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