Making DH10bac competent cells - (Oct/23/2006 )

Hi There,

Does anybody know the protocol for making DH10Bac competent cells? I am thinking of making the competent cells on large scale from DH10Bac cells which are already competent, procured from Invitrogen.

Thanks a lot for any reply you may post.

Here is my prptocol.
These cells can't be electroporated so they are chemically competent cells

Préparation cellules compétentes DH10bac :

Culture ON 1ml in LB+Kan + Tétra
Seed 100ml LB Kan+ Tétra
Grow 37°C agitating til OD reaches OD600nm= 06-08
Pellet cells 3000 rpm 15 min 4°C
Wash cells with same volume of CaCl2 50 mM
Let sit 15' 4 °C, pellet cells 15 min 3000 rpm 4°C
Resuspend carefully in 4 ml CaCl2 + 15% glycérol

Check with 1 µL of miniprep de pFastbac