Protocol Online logo
Top : Forum Archives: : Molecular Biology

Restriction Mapping Problem - (Aug/26/2001 )

Hi Everyone,

I'm trying to produce a high resolution restriction map of a cloned 10kb piece of archaebacterial DNA. I've tried a variety of enzymes on plasmid DNA isolated with a miniprep kit, but I'm constantly getting partial cutting with most of the 20 enzymes I'm using. I've tried different amounts of DNA in the digest, and digested for differing times, but to no avail! It happens even for non-methylation sensitive enzymes.Can anyone offer any advice as to how to prevent partial cutting, otherwise I'm going to pull my hair out!

Thanks a lot!



Hi, Chris!Have you tried with I - CeuI? It cuts in rrna region. If you're lucky, your piece of DNA might contain some repeats...Good luck.