Optimization of DNA hybridization on aminopropyl-controlled pore-glass particles: Detection of non-labeled targets by picogreen staining

摘要

A rapid and cost effective strategy was developed to detect the hybridization of DNA sequences, covalently attached to aminopropyl-controlled pore glass particles (ACPG) with non-labeled targets. The optimization of parameters, such as immobilization chemistry, hybridization time, and washing conditions, was first accomplished using a direct method based on the detection of fluorescein-labeled complementary targets. Under the optimized conditions, a methodology based on the post PicoGreen staining of the duplex molecules formed upon hybridization was demonstrated. This approach eliminates the requirements for pre-target labeling, while providing a rapid detection (less than 20 minutes) of specific complementary targets present in the solution.