Team:HokkaidoU Japan/Notebook/Color R0040-B0034-(K592010, K592012, K864401)-B0015

From 2013.igem.org

Maestro E.coli

Notebook

R0040-B0034-(K592010, K592012, K864401)-B0015

  • Start

  • Get chromoprotein DNA

    Great thanks to iGEM Uppsala!
  • Transformation

    K592010, K592012, K864401
    each of 1µL DNA to DH5α
  • Colony PCR & Culture

    K592010, K592012, K864401, B0015
    We confirmed transformation was completely successful.
    Cultured them for subsequent experiments.
  • Mini-prep

    K592010, K592012, K864401, B0015
  • PCR & PCR purification

    K592010, K592012, K864401
    100b up, 200b dn, KOD Plus NEO
  • Digestion

    Cut B0015 with EcoRI, XbaI (using 10xM buffer)
    Cut K592010, K592012, K864401 with EcoRI, SpeI (using 10xH buffer)
  • Antarctic phosphatase

    B0015
  • Ligation

    Ligate B0015 with K592010, K592012, K864401
  • Transformation

    (K592010, K592012, K864401)-B0015
    each of 5µL DNA to DH5α
  • PCR & PCR purification

    R0040-B0034 from our stock
    100b up, 200b dn, KOD Plus NEO
  • Colony PCR

    (K592010, K592012, K864401)-B0015
  • Culture

    (K592010, K592012, K864401)-B0015
  • Mini-prep

    (K592010, K592012, K864401)-B0015
  • Digestion

    Cut K592010-B0015 with EcoRI, XbaI (using 10xM buffer)
    Cut R0040-B0034 with EcoRI, SpeI (using 10xH buffer)
  • Antarctic phophatase

    (K592010, K592012, K864401)-B0015
  • Ligation

    Ligate (K592010, K592012, K864401)-B0015 with R0040-B0034
  • Transformation

    R0040-B0034-(K592010, K592012, K864401)-B0015
    each of 5µL DNA to DH5α
  • Culture

    R0040-B0034-(K592010, K592012, K864401)-B0015
  • Mini-prep

    R0040-B0034-(K592010, K592012, K864401)-B0015
  • Complete!