Revision as of 15:08, 7 August 2015

Thursday 30th July

Lab Work

Transformation

by Audrey

Biobricks:

• BBa_K1707004
• BBa_K1707009

As usual

Plasmid extraction

by Johan

Biobricks:

• BBa_K1707008 #3, #4, #5 and #6

With the Macherey-Nagel Extraction kit

Ligation

by Pauline

• BBa_K1707011: BBa_B0030 + BBa_K1707010 #1
  • 3 µL BBa_B0030
  • 5 µL BBa_K1707010
  • 2 µL Buffer Ligase 10x
  • 1 µL Ligase

• BBa_K1707021: BBa_K1707000 + BBa_K1707010
  • 1,5µL BBa_K1707000
  • 5 µL BBa_K1707006
  • 1 µL Buffer Ligase 10x
  • 1 µL Ligase
  • 1,5 µL H2O

Incubation 4h, 4°C

Digestion Verification

by Pauline

• Biobricks:
  • BBa_K1707008 #2, #3, #4, #5, #6
  • BBa_K1707005 #1 and #2

• Mix:
  • 1 µL Buffer FastDigest 10x
  • 0,5 µL XbaI
  • 2 µL Plasmid
  • 6,5 µL H2O

Incubation 1h30, 37°C

Electrophoresis

by Pauline

Agarose gel 1%, migration 80V then 110V

We can conclude that BBa_K1707008 isn't OK, and BBa_K1707005 #1 and #2 are OK

New Culture

by Pauline

Biobricks:

• BBa_K1707012
• BBa_K1707013
• BBa_K1707019

4 clones for each, in 5ML LB + 20 ng/µL Chloramphenicol

Transformation

by Johan and Pauline

Biobricks:

• BBa_K1707021
• BBa_K1707011
• BBa_I13600

As usual

Ligation

by Coralie

• BBa_K1707008: BBa_J23101 + BBa_K1707003
  • 2,5 µL BBa_J23101
  • 11 µL BBa_K1707003
  • 2 µL Buffer Ligase 10x
  • 1 µL Ligase
  • 3,5 µL H2O

Incubation over night, 16°C

Soil experiment

by Coralie

Soil

Observation of J0 plates: there are a lot of colony in each. Contamination is alright. Controls (+) and (-) are Ok too. We take 1g of each pot of soil and we dilute it in 5mL sterile H2O. After shaking and decant, we put 100µL of supernatant on the right plate. 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

We use a less powerful adjustment to see tall the result than the day before.

Member present:

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

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