Protocol Online logo
Top : Forum Archives: : Molecular Biology

RFLP electrophoresis - (Mar/12/2006 )

hi all
I am doing RFLP analysis , I need to differentiate between 100 bp-150bp-200bp-220bp restriction fragments ,but with ordinary agarose gel these are indistinguishable.
I will be very thankful for any suggestion for the kind of agarose gel and/or electrophoresis conditions,


You should have no trouble at all distinguishing these lengths. Try a 2% normal agarose gel, TAE, 8 volts/cm for 80 minutes. Run a ladder to debug this problem -- any ladder with 100 bp resolution would work, but we run the NEB 2-log ladder, which can be used for everything.



Other option that works well, specially if you have a lot of bands, is poliacrilamyde gels and silver staining. I once had to made RAPD and used this geles to compare the different patterns and was better than the 2% agarose gel.