Bio-Rad Mini Trans-Blot® Cell - Manual (Page 26)

20 Mini-Trans-Blot Electrophoretic Transfer Cell
Greater retention during sequencing manipulations
enhances the likelihood of obtaining information from
rare, low abundance proteins, by increased initial coupling
and higher repetitive yields. In addition, PVDF membrane
exhibits better binding efficiency of blotted material in the
presence of SDS in the transfer buffer. PVDF must first
be wetted in 100% MeOH but can then be used in buffer,
which does not contain MeOH.
5.2 DNA and RNA Blotting Membranes
Zeta-Probe
® Nylon Membrane
Nitrocellulose is not a suitable medium for electrophoretic
transfer of nucleic acids, as high concentrations of salt
(>10x SSC) are required for efficient binding.
13 Molecules
≤500 bp are not bound at all, even at high salt. Low
resistance results when an electric current is passed
through a solution of high salt. This causes potentially
damaging high currents (and power) even at very low
voltages. Since V/cm is the eluting force, inefficient
transfer occurs under conditions required for proper
binding. Zeta-Probe membrane allows efficient binding of
all sizes of single stranded DNA and RNA in the presence
of low ionic strength buffers.
13 Zeta-Probe membrane
is an ideal alternative to nitrocellulose for the transfer of
nucleic acids. Binding is more stable through post transfer
washes, and reprobing may be performed as many as 10
times.
A variety of blotting membranes is available for
immunoblotting, each with particular advantages
depending on the needs of the experiment. The
physical properties and performance characteristics of a
membrane should be evaluated when selecting the
appropriate transfer conditions.