2 replies to this topic

[silvepa23]

Hi,

I was given an aliquot of invitrogen top 10 competent cells.  I was wondering how to go about growing more. Is it simply a matter of be very careful and putting a small amount on an LB plate without antibiotics.  Since competent cells arent antibiotic resistant how do i know the colonies are actually clones of the top 1o competent cells?  thanks and any help would be greatly appreciated.

[GNANA]

Just streak on a fresh agar plate without antibiotic, parallely you can also streak them in an antibitic plate where you wont get any colonies...thats it, next day you can pick a colony and grow them for competent cell preparation...This wouldnt fully answered your question but this is how i do, since i have my own stock comp cells i am confident that my handling would have not caused any contamination in the stock....

[bob1]

Protocol for Top10 competent cell preparation here.  It works fine in my experience.