I designed 2 dual hybridization probes for my single SNP. where I attach my fluorophore 5'-FAM-nnnn-BHQ1-3', and 5'-HEX-nnnnnn-BHQ2-3'. I need to run it on the MIC RT PCR machine by Biomolecular Systems. when I select the program for dual hybridization probe detection, It gives pre-enlisted quenchers at 5' end and fluorescent dye at 3'nd. can I choose inverse and still get results? or should I use a different machine or a different probe? kindly suggest any troubleshooting. I need to detect two targets in a single vial. can I use eliquates of the same sample no and add Fam probe in one and hex prob in the other and run as a hydrolysis probe setup/ will this approach give results?
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