Methods for the rapid amplification of extremely low quantity nucleic acids in a sample are provided. The disclosed methods are capable of amplifying less than 1 pg of DNA and/or RNA from a biological sample using a single tube and one-step or two-step preparation.

The invention relates to a single novel dry reagent composition for use in subsequent nucleic acid amplification, such as PCR (Polymerase Chain Reaction) and the isothermal LAMP (loop-mediated isothermal amplification), the composition including a nucleic acid amplification enzyme; a corresponding enzyme binder; a sugar-based stabiliser; deoxyNucleotide TriphosPhates; oligonucleotides; and a zwitterionic agent where a tris-based buffer is specifically excluded. The invention further relates to methods for producing said single dry reagent composition using air drying processes.

The invention relates to a single novel dry reagent composition for use in subsequent nucleic acid amplification, such as PCR (Polymerase Chain Reaction) and the isothermal LAMP (loop-mediated isothermal amplification), the composition including a nucleic acid amplification enzyme; a corresponding enzyme binder; a sugar-based stabiliser; deoxyNucleotide TriphosPhates; oligonucleotides; and a zwitterionic agent where a tris-based buffer is specifically excluded. The invention further relates to methods for producing said single dry reagent composition using air drying processes.

Lyophilized biological reagents, such as enzymes (e.g., PCR reagents) and antibodies, are provided that include a wax component. Thus, in some aspects, a method is provided for storing a biological reagent comprising formulating the reagent into a lyophilized composition including a wax component. Methods for using such lyophilized reagents are likewise provided.

Lyophilized biological reagents, such as enzymes (e.g., PCR reagents) and antibodies, are provided that include a wax component. Thus, in some aspects, a method is provided for storing a biological reagent comprising formulating the reagent into a lyophilized composition including a wax component. Methods for using such lyophilized reagents are likewise provided.

The present disclosure provides polynucleotide-based inhibitors for reversible activation of DNA polymerases. Use of lower Tm polynucleotide-based inhibitors allow PCR reaction assembly at room temperature while activating polymerase at higher PCR primer annealing temperatures, where the reversible nature of the inhibition additionally improves priming specificity during each PCR cycle. Additionally, temperature controlled inactivation of polymerase activity after PCR or other polymerase based enzymatic incubation eliminates a purification step when needed for compatibility with subsequent enzymatic incubations. For this application, the T.sub.m of the polynucleotide-based inhibitor is higher than the desired reaction conditions of the subsequent enzymatic incubation.

The present disclosure provides polynucleotide-based inhibitors for reversible activation of DNA polymerases. Use of lower Tm polynucleotide-based inhibitors allow PCR reaction assembly at room temperature while activating polymerase at higher PCR primer annealing temperatures, where the reversible nature of the inhibition additionally improves priming specificity during each PCR cycle. Additionally, temperature controlled inactivation of polymerase activity after PCR or other polymerase based enzymatic incubation eliminates a purification step when needed for compatibility with subsequent enzymatic incubations. For this application, the T.sub.m of the polynucleotide-based inhibitor is higher than the desired reaction conditions of the subsequent enzymatic incubation.

Methods for the rapid amplification of extremely low quantity nucleic acids in a sample are provided. The disclosed methods are capable of amplifying less than 1 pg of DNA and/or RNA from a biological sample using a single tube and one-step or two-step preparation.

Methods for the rapid amplification of extremely low quantity nucleic acids in a sample are provided. The disclosed methods are capable of amplifying less than 1 pg of DNA and/or RNA from a biological sample using a single tube and one-step or two-step preparation.

The disclosure provides materials and methods for detection of pathogens. In particular, the disclosure provides primers and compositions for detection of viral pathogens, such as SARS-CoV-2. In addition, the disclosure provides a warm-start digital CRISPR assay for detection of viral pathogens, including SARS-CoV-2.