Team:Paris Saclay/Notebook/August/12

Wednesday 12th August

Lab Work

Purification

by Pauline

Biobricks:

  • BBa_I13602 #1
  • BBa_I13602 #2
  • BBa_K1707020
  • BBa_K1707030

With Macherey-Nagel Purification kit


Quantification

by Pauline

Agarose gel 1%, Migration 90V

We can conclude:

  • BBa_I13602 #1: 40 µg/µL
  • BBa_I13602 #2: 40 µg/µL
  • BBa_K1707020: 10 µg/µL
  • BBa_K1707030: 13 µg/µL


Digestion

by Coralie

Biobricks:

  • BBa_K1707011 #3 and BBa_K1707012 #2
    • Mix for each reaction:
      • 1 µL PstI
      • 1 µL XbaI
      • 2 µL Buffer FastDigest 10x
      • 6 µL H2O
      • 10 µL plasmid
    • Incubation 37°C, 2h


Purification on gel

by Pauline

Agarose gel 1%, 90V

Biobricks:

  • BBa_K1707011 #3
  • BBa_K1707012 #2

We can conclude that biobricks are OK

Purification by the Macherey-Nagel Purification kit


Ligation

by Pauline

  • BBa_K1707035: BBa_K1707030 + BBa_I13602
    • 11,6 µL BBa_K1707030
    • 1,25 µL BBa_R0040
    • 2 µL Ligase
    • 2 µL Buffer 10x
    • 3,15 µL H2O
  • BBa_K1707036: BBa_K1707020 + BBa_I13602
    • 15,5 µL BBa_K1707006
    • 1,25 µL BBa_B0030
    • 2 µL Ligase
    • 2,5 µL Buffer 10x
    • 3,75 µL H2O


Incubation 4°C, ON


New culture

by Pauline

Biobricks:

  • From stock:
    • BBa_K1707012 #2 x2
    • BBa_K1707011 #3 x2
    • BBa_R0051
  • Two clones from plate of each:
    • BBa_K1707033
    • BBa_K1707031
    • BBa_K1707016

In 5mL LB + 5µL of appropriate antibiotic


Soil experiment

Member present:

  • Instructors: Claire
  • Students: Coralie and Pauline

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