I'm now using a 280bp DIG labelled probe in Southern hybridization to
detect a single copy gene in mammalian genomic DNA. However, the results
were very unsatifactory even 50ug od DNA were loaded into each lane. No
positive bands and results obtained so far. I've followed exactly what
the DIG user manual said, but no help.
And do any one know if dextran sulfate or PEG can be added and use in
DIG Southern hybridization??
Besides, the efficiency of DNA transfer in Southern was also very low,
large amount of DNA left in the gel after the process. I suspect the
problem was due to the use of an excessively heavy weight at the top. Do
anyone knows if it was really the cause..???