Team:Paris Saclay/Notebook/August/6

Thursday 6th August

Lab Work

Soil experiment

by Coralie

Water

Observation of plates (05/08/2015) Almost all plates are yellow. We choose to stop the experiment.

We will define a new protocol:

• Count of bacteria before contamination
• Evaluation of the count of bacteria we should see at Day0
• Measurement of the pH before each sampling (We think that maybe the water pH change with adding the LB)

Low Budget Challenge

by Coralie

We can't say a lot of things... But we can see some colony on the agar with the baker's yeast in the agar. We try another time with different agar recipes:

• Agar 1:
  • 0,27g Stock Cube
  • 0,600g Agar Agar
  • 0,135g Baker's yeast
  • 0,225g sugar
  • 45mL mineral water

• Agar 2:
  • 0,27g Stock Cube
  • 0,600g Agar Agar
  • 0,135g Baker's yeast
  • 45mL mineral water

We make striations with BBa_J23101+BBa_I13504 strain with a curved pin. We do the same on an usual LB plate without antibiotics (Control +)

Incubation over night, 37°C

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

Member present:

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

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