Analytical standards can allow one to detect and/or measure sampling, processing, and/or amplification errors in a sample that includes a plurality of polynucleotide molecules. The analytical standards can provide an internal control to detect errors in the representation of the original sample reflected in data obtained after manipulating and/or processing of sample molecules.

Analytical standards can allow one to detect and/or measure sampling, processing, and/or amplification errors in a sample that includes a plurality of polynucleotide molecules. The analytical standards can provide an internal control to detect errors in the representation of the original sample reflected in data obtained after manipulating and/or processing of sample molecules.

In a method for comparative analysis, the expression levels of the target miRNAs in each body fluid sample are corrected using the expression level(s) of a correcting endogenous miRNA(s) that is/are simultaneously measured with the expression levels of the target miRNAs in the sample. As the correcting endogenous miRNA(s), one or more miRNAs selected from specific 10 kinds of correcting endogenous miRNAs is/are used. Comparative analysis of target miRNAs among body fluid samples can be carried out more accurately than by conventional techniques.

In a method for comparative analysis, the expression levels of the target miRNAs in each body fluid sample are corrected using the expression level(s) of a correcting endogenous miRNA(s) that is/are simultaneously measured with the expression levels of the target miRNAs in the sample. As the correcting endogenous miRNA(s), one or more miRNAs selected from specific 10 kinds of correcting endogenous miRNAs is/are used. Comparative analysis of target miRNAs among body fluid samples can be carried out more accurately than by conventional techniques.

PCR that allows the researchers to amplify a desired DNA requiring only tiny amounts of sample. Such amplification reactions are technically challenging and are often hampered by several practical issues such as the presence of PCR inhibitors, sample degradation and low quantities of said sample. The invention addresses these issues with a method for evaluating the amplification efficiency and/or the presence of inhibitors and/or degradation and/or performing a quantification of a nucleic acid in a real-time amplification reaction comprising: optionally amplifying in a reaction composition a first target nucleic acid using a first primer pair in a real-time amplification reaction, (i) amplifying in said reaction composition one or more second internal nucleic acid control templates (IC) with a length of between 50 and 2000 nucleotides, wherein the second nucleic acid has a sequence selected from the group of: i) SEQ ID NO. 1 or a sequence that is differs by no more than 15% therefrom, ii) a reverse complement of SEQ ID NO. 1 or a sequence that is differs by no more than 15% therefrom, iii) a sequence or a reverse complement thereof comprising the final 18 to 30 3′-nucleotides of SEQ ID NO. 1 at its terminal 3′-end and the final 18 to 30 5′-nucleotides of SEQ ID NO. 1 at its terminal 5′-end, or terminal ends that differ by no more than 5% from SEQ ID NO. 1, wherein between these terminal 3′-ends and 5′-ends the nucleic may have any nucleotide sequence and is between about 30 and about 1950 nucleotides in length.

Provided herein is technology relating compositions and methods for analysis of methylated DNA from a subject. The technology also relates to use of endogenous methylated DNAs as internal controls for marker gene methylation assays.

Provided herein is technology relating compositions and methods for analysis of methylated DNA from a subject. The technology also relates to use of endogenous methylated DNAs as internal controls for marker gene methylation assays.

Embodiments of a method and/or system for facilitating characterization of one or more conditions can include: generating a set of target-associated molecules; generating a reference-associated set of molecule; facilitating generation of at least one spike-in mixture; determining one or more abundance metrics based on an analysis of the at least one spike-in mixture; and facilitating the characterization of the one or more conditions based on the one or more abundance metrics.

Embodiments of a method and/or system for facilitating characterization of one or more conditions can include: generating a set of target-associated molecules; generating a reference-associated set of molecule; facilitating generation of at least one spike-in mixture; determining one or more abundance metrics based on an analysis of the at least one spike-in mixture; and facilitating the characterization of the one or more conditions based on the one or more abundance metrics.

Among other things, a method for normalizing a sample is provided. In some embodiments, the method comprises: (a) reacting a sample with a limiting amount of a single-turnover sequence-specific endonuclease that recognizes a target sequence, thereby cleaving a portion of the nucleic acid molecules that comprise the target sequence and producing a normalized amount of a first cleavage product; and (b) isolating, transcribing or selectively amplifying the normalized amount of the first cleavage product. In this method, because a limiting amount of the endonuclease is used, the normalized amount of the first cleavage product is determined by the limiting amount of the first single-turnover sequence-specific endonuclease used in step (a).