Team:Paris Saclay/Notebook/July/24


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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