E Coli strain for lentiviral packing vector
Posted 17 July 2011 - 01:03 AM
I've got some lenti vectors but I can't get the packaging vector (delta R) to transform into my dh5alpha. I only bought the cheapest subcloning efficiency ones (they worked for all the others, including the supposedly toxic insert....
I've tried transforming 2ng, 5ng and 10ng but nothing. I am going to plate the rest of the bacteria but I suspect there won't be much there.
We have some TOP10 in the lab, but can anyone recommend another strain to transform something over 10kb?
Posted 17 July 2011 - 01:45 PM
Maybe you should also increase the amout of DNA you use when working with large plasmids.
Posted 19 July 2011 - 11:50 PM
Guess I will have to buy the STBL lines then
Posted 20 July 2011 - 02:15 AM
Posted 20 July 2011 - 05:11 AM
we don't use ethidium so will have to run at least 50ng.