restriction digestion - (Nov/27/2004 )
i'm not sure if any of you are familiar with restriction enzyme Fse1. I have been having a lot of trouble with this enzyme because it doesn't always cleave my plasmid; it's also a really expensive enzyme. Does anyone have any background information on this RE? what is the optimal concentration of this enzyme i should use?
are you sure that your plasmid has got FseI site ?you have told that you are getting problems in restriction. are u getting some sort of band on the gel after digestion? if so try to screen for the purity of the DNA. let me know if u get any banding pattern,
Check you have dissolved the DNA in water instead of TE buffer where EDTA interfere with restriction digestion.
Also prolong the incubation time
Hope you are taking sufficient enzyme for the digestion, pls check it alos