Abstract: The present disclosure relates to particle-based methods and kits for extracting nucleic acids, such as with magnetic or magnetically-responsive particles, from a wide variety of biological samples. The methods and kits of this disclosure may be used to extract nucleic acids with high purity and recovery (compared to conventional methods, such as using spin-columns) from simple through complex biological samples. The methods and kits of this disclosure may be used in high-throughput extractions or may be amenable to use in high-throughput applications.

Abstract: Assays using enzymatic degradation of RNA/DNA heteroduplexes are provided for the detection of target nucleic acid molecules. Such enzymatic degradation may be obtained by enzymes including RNaseH. Exemplary methods include Probe Trapping (PT), Hybrid Energy Transfer (HET), and Fluorescent Probe Degradation (FPD). The assays make use of an RNA or chimeric RNA/DNA probe which recognizes a target DNA sequence and forms an RNA/DNA heteroduplex that is the substrate for the enzyme RNaseH which degrades the RNA portion of the probe. Degraded probe fragments diffuse away from the DNA target leading to a detectable signal and allowing the DNA target to hybridize to another probe. Probe degradation is cycled over time. The assays disclosed herein are suitable for use in detecting DNA sequences and could be used in a medical diagnostic.