Team:Paris Saclay/Notebook/July/8

Wednesday 8th July

Lab Work

Transformation

by Audrey

Biobrick: R0051 (2015) On LB + Ampicillin 100ug/mL

Rehydratation

by Seong Koo, Johan

R0051 from different years (2013 and 2014)

Digestion

by Pauline, Coralie

First Digestion:

  • Biobricks:
    • J23101
    • J23106
    • J23117
  • Mix:
    • 10µL of our plasmid with promotor
    • 1µL SpeI
    • 1µL PstI
    • 2µL buffer 10x FastDigest
    • 6µL H2O

Second Digestion:

  • Biobricks:
    • K115017
    • I13504
  • Mix:
    • 10µL of our plasmid with gene
    • 1µL XbaI
    • 1µL PstI
    • 2µL buffer 10x FastDigest

After this step, we separate the sequence we need from the sequence we don't with electrophoresis


Electrophoresis

by Johan, Coralie, Seong Koo, Pauline

Preparation of Agarose Gel 1%, 0,5g in 50mL of 1X TAE, 0,5uL of BET Migration 0,06A 80V

  • K115017
  • I13504

Ligation

by Coralie, Audrey

  • J23117 + I13504
    • 2,5µL J23117
    • 15µL I13504
    • 1µL T4 DNA Ligase
    • 2µL Buffer T4 DNA Ligase 10x
  • J23106 + I13504
    • 1,5µL J23106
    • 15,5µL I13504
    • 1µL TA DNA Ligase
    • 2µL Buffer T4 DNA Ligase 10x
  • J23201 + I13504
    • 2µL J23101
    • 14,5µL I13504
    • 1µL Ligase
    • 2µL buffer 10x

Incubation ON, 16°C

Members present:

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