Team:HokkaidoU Japan/Notebook/Pre experiment/Stem replacement

From 2014.igem.org

Revision as of 11:26, 14 October 2014 by Osamu (Talk | contribs)

Stem Region Replacement

  • Start

  • PCR

    pHN1257 161 Tm: 162 Tm: KOD FX NEO

  • Transformation

    R0010 on ice 30min each of 5.0µL DNA to DH5α 42°C 1min on ice 2min

  • Digestion

    Cut R0010 with PstI, SpeI (using 10xH buffer) and pHN1257 with XbaI, PstI (using 10xM buffer)

  • Ligation

    Ligate R0010 with pHN1257

  • Transformation

    Ligation products on ice 30min each of 5.0µL DNA to DH5α 42°C 1min on ice 2min

  • Colony PCR

    R0010-stem-dT 100up f 162primer R(あとで書く)

  • mini-Prep

    R0010-stem-dT

  • PCR(confirm)

    R0010-stem-dT 100up 162 KOD FX NEO

  • Complete!

Retrieved from "http://2014.igem.org/Team:HokkaidoU_Japan/Notebook/Pre_experiment/Stem_replacement"