Team:Bordeaux/NotebookJune17

From 2014.igem.org

17th June

Molecular biology

  • Plasmid extraction of VPGXG-20,-40 and VPGXG-60 in pSB6A1 by QIAprep Spin Miniprep Kit from QIAGEN


Production

  • A600 preculture VPGXG-20 = 1,994
    A600 preculture VPGXG-40 = 1,849
    A600 preculture VPGXG-60 = 2,177
    - Culture of VPGXG-20, VPGXG-40 and VPGXG-60 at 37°C

    T=0, A600 VPGXG-20 = 0,037
    A600 VPGXG-40 = 0,016
    A600 VPGXG-60 = 0,018
    - IPTG induction when A600= 0,8 and culture at 25°C

    T=8h, 600 VPGXG-20 = 1,845
    A600 VPGXG-40 = 1,509
    A600 VPGXG-60 = 1,499


Purification

  • We stop protocol after resuspend cell pellet in 20mL PBS and store at -80°C.


Design

  • Pattern : Met-M-[(VPGGV)2VPGQQVPGGY]-Spe1-Stop
  • We try to design oligo with this pattern