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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?
3rd Oct, 2012
Mohan Harikrishnan
Louisiana State University
Before going for hybridization check the quality and concentration of your probe. If you know the sequence details of your target gene along with UTR region, then you can use 3'UTR region as probe for hybridization. If you are using kit based detection check the troubleshooting guide to solve your problems.