Folks, We want to amplify two different fragments by PCR and detect them by flow cytomety. For the first fragment, we can use a primer marked by rhodamine green and for the second one we need to emit in the red. So a possible solution is the FRET technique. My question is: What constraints do one has to face when designing the primers...(Distance between the two fluorescent markers, compatibility of the markers, can both markers be bound on the same primer...) Thanks in advance for input. Olivier Lesueur Unilever Research, NL
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