Improved chemiluminescent DNA sequencing.

A new chemiluminescent 1,2-dioxetane substrate, CSPD, shows improved performance over AMPPD when used in our nonisotopic method for DNA sequencing. CSPD differs from AMPPD by the addition of a chlorine atom to the adamantyl group that limits the amount of aggregation of the dioxetane and its dephosphorylated anion. This results in a shorter time elapsing before reaching steady state light emission when detecting nucleic acids on nylon membrane. An additional advantage of CSPD over AMPPD is that the resolution of imaged DNA bands does not degrade over time. These features of CSPD permit rapid acquisition of high-quality DNA sequence data.