CHEMILUMINESCENT SUBSTRATES FOR DETECTION OF RESTRICTION-FRAGMENT-LENGTH-POLYMORPHISM

Five new enzyme-triggered dioxetane substrates were evaluated for rest riction fragment length polymorphism (RFLP) analysis of HaeIII-restric ted DNA. Of these, one substrate designated CDP-Star provided unsurpas sed sensitivity within one working day without the presence of an enha ncer. Far greater sensitivity was obtained from chemiluminescent detec tion of DNA on MSI neutral membranes than the sensitivity obtained fro m six day film exposures of P-32 labelled insert probes on PALL B memb ranes, including the detection of most low molecular weight alleles. F or nylon membranes better suited for alkaline phosphatase-triggered ch emiluminescent detection of DNA, high salt/neutral pH southern transfe r conditions were better than alkaline southern transfer conditions.