Chemiluminescent substrates for detection of restriction fragment length polymorphism.

Five new enzyme-triggered dioxetane substrates were evaluated for restriction fragment length polymorphism (RFLP) analysis of HaeIII- restricted DNA. Of these, one substrate designated CDP-Star provided unsurpassed sensitivity within one working day without the presence of an enhancer. Far greater sensitivity was obtained from chemiluminescent detection of DNA on MSI neutral membranes than the sensitivity obtained from six day film exposures of 32P labelled insert probes on PALL B membranes, including the detection of most low molecular weight alleles. For nylon membranes better suited for alkaline phosphatase-triggered chemiluminescent detection of DNA, high salt/neutral pH southern transfer conditions were better than alkaline southern transfer conditions.