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Asked 3rd Oct, 2012
  • Central Proteina Prima Company of Jakarta, Indonesia

How to design probe specific hybridize with a specific targeted gene in Southern blot method?

I use DIG-11-dUTP label probe with specific targeted gene to detect in host genome. Host genome have a huge sequence that to date no information in data base. I cut the genomic DNA by restriction enzymes but after southern blot process my specific gene in the host genome does not show the signal band. I use chemilluminence (SDP star) substrate for alkaline phospatase, I do not use radioactive label 32P. Could anyone give me more ideas with this case?
4th Oct, 2012
Heny Budi Utari
Central Proteina Prima Company of Jakarta, Indonesia
I have checked the probe already, and I also have done for probe detection with different concentration, I used the high concentration that show positive signal. My restriction enzyme digested genomic DNA did not show a signal but my positive undigested plasmid show the band ( I load 20 ng/ul of plasmid positive)....could you give any suggestion to me...