large DNA plasmid prep

[plxsrt at pln1.nott.ac.uk]

Lakshman Ramamurthy PhD <lr50916 at glaxowellcome.com> wrote:

>Netters:

>I have tried unsuccessfully to make a Qiagen large prep of a 23 kb
>plasmid DNA.. Do you have any suggestions to get around this problem. I
>need about 100 micrograms of DNA
>Thanks
>LR

The bit in the manual about using larger volumes of culture seems to
be critical for large plasmids/cosmids.  I'd check how well your
plasmid grows using a crude prep before using the Qiegen.  If it
doesn't grow well then bump up the culture volume.



Simon T