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