Inoue again....

Paul doesburg at pa1.fgg.EUR.NL
Fri Jun 14 08:29:03 EST 1996


HI,
I've tried making competent cells according to the Inoue protocol but 
just can't reach a competence of 10^8, let alone 10^9, with either 
DH5alpha or XL2BLUE cells(stratagenes ultracompetent cellstrain). I seem 
to be stuck at around 7*10^7. My reference plasmid is from a commercial 
source so I assumed I must be doing something else wrong. Now I remember 
the question being asked, but couldn't find a reply whether freezing and 
storing the cells at -80 instead of N2 decreases the competence. And 
is LB soft agar really needed?



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