Bio-Rad Mini Trans-Blot® Cell - Manual (Page 24)

18 Mini-Trans-Blot Electrophoretic Transfer Cell
4.2 Optimizing DNA and RNA Transfer
Problems with elution of nucleic acids can be solved by
altering the gel percentage. It may be somewhat more
difficult to quantitatively transfer large amounts of DNA
used in genomic blots. Agarose gels over 6 mm thick are
not compatible with the Mini Trans-Blot. The following
tactics should be considered for optimizing elution in such
transfers.
Alter gel composition
Lower % total monomer or % crosslinker for
polyacrylamide gels.
Lower % agarose. This allows better elution of high
molecular weight DNA.
Alter DNA denaturants
It has been found that glyoxal denaturation allows more
efficient elution of DNA than NaOH. Boiling polyacrylamide
gels to denature DNA has also been found to give
excellent results.
12 Base denaturation often causes
polyacrylamide gels to weaken and stick to blotting
membranes.