Abstract: The invention provides methods of digital amplification using primers of limited nucleotide composition. Limited nucleotide composition means that the primers are underrepresented in at least one nucleotide type. Such primers have much reduced capacity to prime from each other or to extend initiated by mispriming from other than at their intended primer binding sites in a target nucleic acid.

Abstract: The invention provides methods of amplification from a single primer or a pair of forward and reverse primers of limited nucleotide composition. Limited nucleotide composition means that the primers are underrepresented in at least one nucleotide type. Such primers have much reduced capacity to prime from each other or to extend initiated by mispriming from other than at their intended primer binding sites in a target nucleic acid.

Abstract: Various embodiments are directed to a fluorescence reader for assessing a fluorescence intensity of one or more substance samples. The fluorescence reader may comprise a sample support structure for accepting sample wells containing corresponding samples therein, an illumination structure positioned below the sample support structure and configured to illuminate one or more samples positioned within the sample wells, and an imaging structure positioned above the sample support structure and configured to detect a fluorescence emitted by the one or more samples. The illumination structure may comprise one or more reflective perimeter walls, together with the support structure, defining an enclosed illumination cavity, and one or more light sources positioned within the enclosed illumination cavity and configured to emit light causing the one or more samples to fluoresce.

Abstract: The invention provides methods of digital amplification using primers of limited nucleotide composition. Limited nucleotide composition means that the primers are underrepresented in at least one nucleotide type. Such primers have much reduced capacity to prime from each other or to extend initiated by mispriming from other than at their intended primer binding sites in a target nucleic acid.