"
Team:Bordeaux/NotebookJune12
From 2014.igem.org
6th June
Molecular biology
- Making competent cell: day 3
- Digestion, ligation and transformation of pSB1C3 and pSB6A1 with our inserts VPGXG-20, VPGXG-40 and VPGXG-60.
Production
A600 preculture VPGXG-20 = 2,349
A600 preculture VPGXG-40 = 2,431
A600 preculture VPGXG-60 = 2,143
T=0, A600 VPGXG-20 = 0,003 A600 VPGXG-40 = 0,005 A600 VPGXG-60 = 0,047
- Culture of VPGXG-20, VPGXG-40 and VPGXG-60 at 37°C
T=0, A600 VPGXG-20 = 0,003 A600 VPGXG-40 = 0,005 A600 VPGXG-60 = 0,047
- IPTG induction when A600 = 0,8 and culture at 25°C
Purification
- We stop protocol after resuspend cell pellet in 20mL PBS and store at -80°C.
