if i designed a pair of primers and examined their specificity to the exact gene and by PCR i obtained the exact band. can i use this antisense primer as oligonucleotide probe directly for performing fluorescent in situ hybridization or i should make gene cloning and sequencing before using it.
Frederic Lepretre
Hi Mona
I would suggest you to be sure first that your primers amplify only your targeted gene. try an in silico PCR to analyze that point (http://genome.ucsc.edu/cgi-bin/hgPcr). at the time this has been done, I would use the amplicon as a probe, not only one of the primer. Your probe will therefore be bigger and the fluorescence intensity too for FISH. Labelled dNTP are available to do that.
fred
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