The present disclosure, at least in part, relates to an engineered RNA (e.g., microRNA and sgRNA), in the absence of an input signal, that is engineered to have a large enough energy gap between the formations of a first secondary structure, which is unrecognizable by an actuator, and a second secondary structure, which is recognizable by an actuator (e.g., Drosha and Cas protein).

The present disclosure, at least in part, relates to an engineered RNA (e.g., microRNA and sgRNA), in the absence of an input signal, that is engineered to have a large enough energy gap between the formations of a first secondary structure, which is unrecognizable by an actuator, and a second secondary structure, which is recognizable by an actuator (e.g., Drosha and Cas protein).

A method for amplification of nucleic acids in which substantially use is made of the fact that a pre-defined nucleic acid chain (target sequence) can be multiplied/amplified in the presence of a target sequence-specific activator oligonucleotide. The target sequence-specific activator oligonucleotide causes the separation of re-synthesized complementary primer extension products by strand displacement, so that a new primer oligonucleotide can attach to the respective template strand. The thus formed complex of a primer oligonucleotide and a template strand can initiate a new primer extension reaction. The thus formed primer extension products in turn function as templates, so that an exponential amplification reaction results. Amplification of a particular target sequence takes place more efficiently in case of perfect match complementary base pair formation between the activator oligonucleotide and the corresponding target sequence. Mismatches between the activator oligonucleotide and a particular target sequence can result in less efficient amplification. The efficiency of synthesis of perfect match target sequences and mismatch sequences can be measured and compared.

A method for amplification of nucleic acids in which substantially use is made of the fact that a pre-defined nucleic acid chain (target sequence) can be multiplied/amplified in the presence of a target sequence-specific activator oligonucleotide. The target sequence-specific activator oligonucleotide causes the separation of re-synthesized complementary primer extension products by strand displacement, so that a new primer oligonucleotide can attach to the respective template strand. The thus formed complex of a primer oligonucleotide and a template strand can initiate a new primer extension reaction. The thus formed primer extension products in turn function as templates, so that an exponential amplification reaction results. Amplification of a particular target sequence takes place more efficiently in case of perfect match complementary base pair formation between the activator oligonucleotide and the corresponding target sequence. Mismatches between the activator oligonucleotide and a particular target sequence can result in less efficient amplification. The efficiency of synthesis of perfect match target sequences and mismatch sequences can be measured and compared.

Provided herein are primers and primer systems having improved specificity and kinetics over existing primers, and methods of use thereof.

Provided herein are primers and primer systems having improved specificity and kinetics over existing primers, and methods of use thereof.

A DNA nano device (2) and a method of producing a DNA nano device (2) for detecting a biomarker are provided. The DNA nano device (2) includes an ensemble of DNA formations. The formations include a detector (4) adapted to accept a target biomarker and thereby to release a trigger DNA sequence; an amplifier (6) adapted for hybridisation with the trigger DNA sequence which thereby releases a key DNA sequence and the trigger DNA sequence or a further trigger DNA sequence; and a responder (8) adapted for hybridisation with the key DNA sequence and thereby to produce a signal. The DNA nano device (2) thereby produces a signal detectable to an observer in response to the detection of the target biomarker.

A DNA nano device (2) and a method of producing a DNA nano device (2) for detecting a biomarker are provided. The DNA nano device (2) includes an ensemble of DNA formations. The formations include a detector (4) adapted to accept a target biomarker and thereby to release a trigger DNA sequence; an amplifier (6) adapted for hybridisation with the trigger DNA sequence which thereby releases a key DNA sequence and the trigger DNA sequence or a further trigger DNA sequence; and a responder (8) adapted for hybridisation with the key DNA sequence and thereby to produce a signal. The DNA nano device (2) thereby produces a signal detectable to an observer in response to the detection of the target biomarker.

The invention provides compositions and methods for making closed nucleic acid structures in which one or both strands are continuous. The closed nucleic acid structures can be used as sequencing templates among other applications.

The invention provides compositions and methods for making closed nucleic acid structures in which one or both strands are continuous. The closed nucleic acid structures can be used as sequencing templates among other applications.