Team:HokkaidoU Japan/Notebook/RBS SD2,4,6,8-3K3-Replacement

From 2013.igem.org

Maestro E.coli

Notebook

SD2, 4, 6, 8-3K3 Replacement

  • Start

  • PCR

    R-0040-SD(2,4,6,8)-LacZα-B0015 100up 200dn
  • Digestion

    R-0040-SD(2,4,6,8)-LacZα-B0015 EcoRI and PstI (uning 10×H buffer)
    pSB3K3 vector EcoRI and PstI (using 10×H buffer)
  • Antarctic phophatase

    pSB3K3 vector
  • Ligation

    R-0040-SD(2,4,6,8)-LacZα-B0015 with pSB3K3
  • Transformation

    R0040-SD(6/4/2)-LacZα-B0015
    each of 1µL DNA to DH5α
  • Mini-prep

    R-0040-SD(2,4,6,8)-LacZα-B0015 on pSB3K3
  • Sequence

    R-0040-SD(2,4,6,8)-LacZα-B0015 on pSB3K3
  • β-galactosidase assay (1)

    R0040-RBS(SD2/4/6/8)-LacZα-B0015, X/S (as N. C.)
  • β-galactosidase assay (2)

    R0040-RBS(B0034/SD8)-LacZα-B0015, X/S (as N. C.)
  • β-galactosidase assay (3)

    R0040-RBS(B0034/SD8)-LacZα-B0015, X/S (as N. C.)
  • Complete!