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: FACS

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 Cl- and 2 washes with H2O

Member present:

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

Back to the calendar

Team:Paris Saclay/Notebook/July/24

Contents

Friday 24th July

Lab Work

Digestion

Purification

Purification

Quantification

Transformation

Soil experiment

Interlab study: FACS