Abstract: The present invention relates to Multicomponent Nucleic Acid Enzymes (MNAzymes) and methods for their use. MNAzymes comprise two or more oligonucleotide components which self-assemble in the presence of one or more MNAzyme assembly facilitator molecules to form a catalytically active structure. Compositions for making MNAzymes, and collections of MNAzymes are provided. Also provided are methods for using MNAzymes for the detection, identification and/or quantification of one or more targets. The methods can be practiced in solution-based assays or in assays where one or more reaction components are attached to a support structure. The methods allow for multiplexing the MNAzyme detection to detect multiple targets in a single reaction. Also provided are kits for making the compositions, and for practicing the methods provided herein.

Abstract: Provided herein are methods for the combined amplification and detection of one or a plurality of target nucleic acid molecules. The methods encompass the use of an antisense strand of a catalytic nucleic acid in a primer for amplification such that an amplicon produced thereby includes an active catalytic nucleic acid capable of indicating the presence of the target sequences through the modification of a reporter substrate. Devices and kits are also provided. DNA molecules for practicing the methods are also provided. The DNA molecules comprise at least a first portion complementary to at least a first portion of a target nucleic acid sequence, a second portion complementary to an antisense sequence of a second portion of the target nucleic acid sequence, and a third portion comprising an antisense sequence of a catalytic nucleic acid; the third portion positioned between the first and second portions of said DNA molecule.

Abstract: There is described a method for detecting alkylated cytosine in double stranded DNA employing one or more enzymes that differentially modify alkylated cytosine and cytosine. At least one region of the DNA is converted to single stranded DNA and the enzyme is reacted with a target region in the single stranded DNA. The presence or level of alkylated cytosine in the target region is detected by determining the level of enzymatic modification of the target region by the enzyme.

Abstract: The present invention provides a method of detecting a genetic polymorphism in an individual or between individuals. The method comprises the following steps, (1) obtaining a sample containing nucleic acid from an individual; (2) contacting the sample, under conditions which permit primer-initiated nucleic acid amplification and nucleic acid cleavage, with (i) a primer suitable for initiating amplification, (ii) an indicator system which provides a signal proportional to the amount of amplification product, and (iii) a sequence specific nucleic acid cleavage agent; and (3) measuring the signal produced by the indicator system against time. Cleavage of the amplification product by the cleavage agent results in an inhibition of the rate of accumulation of amplification product comprising the sequence recognised by the cleavage agent relative to the rate of accumulation of amplification product not comprising the sequence recognised by the cleavage agent.