PstI digest failure

"sally francis IACR-BB sally.francis at bbsrc.ac.uk
Mon Jul 30 08:20:52 EST 2001


Dear All,

Has anyone ever had problems cutting plant genomic DNA with PstI? The DNA
looks OK when run on a gel, and it can be cut with other enzymes (e.g.
EcoRI) OK. I have tried using a massive amount of PstI (20U) to cut only
400ng in a 50ul reaction, but it still doesn't cut. I tried long reaction
times (24h) - no success. Other people's batches of enzyme from the same
manufacturer (NEB) won't work and neither will PstI from another
manufacturer (Promega). My own PstI cuts plasmids OK, by the way. I can't
use another enzyme for my work, so any helpful suggestions would be greatly
appreciated!

Thanks,

S. Francis 

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