Team:Paris Saclay/Notebook/July/31

Friday 31st July

Lab Work

Plasmid extraction

by Seong-Koo

Biobricks:

  • BBa_K1707012 #1, #2, #3 and #4
  • BBa_K1707013 #1, #2, #3 and #4
  • BBa_K1707019 #1, #2, #3 and #4

With the Macherey-Nagel Extraction kit


Rehydratation

by Pauline

Biobrick: BBa_I13600 (2014-3-19C and 2013-3-20C)


Transformation

by Pauline

Biobricks:

  • BBa_I13600 (2013, 2014 and 2015)
  • BBa_K1707008

As usual

Plasmid extraction

by Seong-Koo

Biobricks:

  • BBa_E0422 #1 and #2
  • BBa_E0022 #1 and #2

With the Macherey-Nagel Extraction kit

New culture

by Pauline

Biobricks:

  • BBa_K1707004: amicillin
  • BBa_K1707011
  • BBa_K1707021

6 clones of each in 5 mL LB

Digestion Verification

by Coralie

  • Biobricks:
    • BBa_K1707012 #1, #2, #3 and #4
    • BBa_K1707013 #1, #2, #3 and #4
    • BBa_K1707019 #1, #2, #3 and #4
  • Mix:
    • 1 µL Buffer FastDigest 10x
    • 0,5 µL XbaI
    • 0,5 µL PstI
    • 2 µL Plasmid
    • 6 µL H2O

Incubation 2h, 37°C

Electrophoresis

by Coralie

Agarose gel, 1% Migration 100V

We can conclude that all biobricks are OK

Soil experiment

by Coralie and Johan

Soil

Observation of J0 plates: there are a lot of colony in each.

Control (-):

  • MCK + Tet: 40 pink colonies
  • MCK + Cm: 10 pink colonies
  • MCK + Spectino: 1 pink colony
  • MCK: 100 pink colonies and a lot of white colonies

We can suppose that we have a contamination between our experiment and control. Or maybe we have a MCK problem because we change it during the experiment.

So, we decide to stop the experiment and do some test:

  • Soil + H2O on new MCK
  • Soil + LB on new MCK
  • LB on new MCK
  • MCK alone

Incubation over night, 37°C


New culture

by Pauline

Biobricks:

  • BBa_K1707005
  • BBa_I13602

A plasmid: PIJ773

Incubation at 37°C, 250 rpm


Member present:

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

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