Friday 24th July

Lab Work

Digestion

by Pauline

Biobricks:

• BBa_K1707003 #4
• BBa_K1399005
• BBa_B0015
• BBa_J23101

In each tube:

• 10 µL plasmid
• 1µL of each enzyme
• 2 µL Buffer FastDigest
• 6µL H2O

Digestions:

• BBa_K1707003: XbaI + PstI
• BBa_K1399005: EcoRI + SpeI
• BBa_B0015: EcoRI + XbaI
• BBa_J23101: SpeI + PstI

Purification

by Pauline

Biobricks:

• BBa_K1707003
• BBa_K1399005

Agarose gel 1%, 110V

We cut bands with a scalpel

Purification

by Audrey

Biobricks:

• BBa_K1707003
• BBa_K1399005
• BBa_B0015
• BBa_J23101

With PCR Clean-up/Gel extraction kit from Macherey-Nigel

Quantification

Biobricks:

• BBa_K1707003
• BBa_K1399005
• BBa_B0015
• BBa_J23101

Agarose gel 1% Migration 110V

Transformation

by Seong-Koo

Biobricks:

• BBa_K1707005
• BBa_K1707006
• BBa_K1707007

Soil experiment

by Seong Koo

Plates observation and count of CFU:

• Strain 1696:
  • Dilution 1: 424 CFU
  • Dilution 0.1: 75 CFU
  • Dilution 0.01: 1 CFU

• Strain 1693:
  • Dilution 1: 524 CFU
  • Dilution 0.1: 35 CFU
  • Dilution 0.01: 0 CFU

• Strain 1320:
  • Dilution 1: 111 CFU
  • Dilution 0.1: 13 CFU
  • Dilution 0.01: 0 CFU

We take 1g of contaminated soil and treat it like yesterday to spread it on specific plates Incubation ON, 37°C

Interlab study: Cytometer

by Johan

We count 500 000 events Controls:

• Cells alones
• Cells transformed by BBa_J23101, BBa_J23106 and BBa_J23117

Our measurements: Cells transformed by

• BBa_J23101 + BBa_I13504
• BBa_J23106 + BBa_I13504
• BBa_J23117 + BBa_I13504

We uses cells in growth phase and stationary phase

Between each test, we do 2 washes with bleach and 2 washes with H2O

Member present:

• Instructors: Alice
• Students: Coralie, Audrey, Pauline, Johan and Seong-Koo

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