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Problem Possible cause
2. Concentrated
probe has contacted
membrane
directly during
probe addition
3. Probe concentration
is too high
4. Probe not denatured
Remedy
2. It is suggested
that up to 1.0 ml
of the buffer used
for prehybridization
is withdrawn for
mixing with the probe.
The mixture should
then be added back
to the hybridization
container in an area
away from the filter.
3. Ensure measurement
of template DNA
concentration is
accurate
4. Non-denatured
double-stranded
probes often
give high backgrounds.