The present disclosure provides, in some embodiments, methods and compositions for exponential amplification of single- and double-stranded DNA under isothermal conditions.

The present disclosure provides, in some embodiments, methods and compositions for exponential amplification of single- and double-stranded DNA under isothermal conditions.

The present application relates to methods for identifying and/or quantifying low abundance, nucleotide base mutations, insertions, deletions, translocations, splice variants, miRNA variants, alternative transcripts, alternative start sites, alternative coding sequences, alternative non-coding sequences, alternative splicings, exon insertions, exon deletions, intron insertions, or other rearrangement at the genome level and/or methylated or hydroxymethylated nucleotide bases, as well as markers to identify early cancer, monitor cancer treatment, and identify early cancer recurrence.

The present application relates to methods for identifying and/or quantifying low abundance, nucleotide base mutations, insertions, deletions, translocations, splice variants, miRNA variants, alternative transcripts, alternative start sites, alternative coding sequences, alternative non-coding sequences, alternative splicings, exon insertions, exon deletions, intron insertions, or other rearrangement at the genome level and/or methylated or hydroxymethylated nucleotide bases, as well as markers to identify early cancer, monitor cancer treatment, and identify early cancer recurrence.

Nucleic acid constructs and methods that provide superior prevention of primer-dimers and other artifacts of false priming events are disclosed. In particular, there is disclosed a hairpin primer having a target-specific primer region, wherein the target-specific region comprises a target-binding dependent cleavage sequence; a first stem forming region 5′ of the target-specific primer region; and a second stem forming region 3′ of the target-specific primer region, wherein the second stem forming region is complementary to the first stem forming region. Methods of using the hairpin primer to amplify a target nucleic acid are also disclosed.

Nucleic acid constructs and methods that provide superior prevention of primer-dimers and other artifacts of false priming events are disclosed. In particular, there is disclosed a hairpin primer having a target-specific primer region, wherein the target-specific region comprises a target-binding dependent cleavage sequence; a first stem forming region 5′ of the target-specific primer region; and a second stem forming region 3′ of the target-specific primer region, wherein the second stem forming region is complementary to the first stem forming region. Methods of using the hairpin primer to amplify a target nucleic acid are also disclosed.

Systems and methods for isothermal amplification of nucleic acids in a ratiometric manner are disclosed. Methods of isothermal amplification of nucleic acids are convenient for areas that lack reliable electricity and/or funding for precision lab equipment. However, these methods typically result in non-ratiometric amplification of target sequences, thus preventing quantitative analysis or application of the results, such as for diagnostic testing. Systems and methods herein provide a method of isothermal amplification that ratiometrically amplifies target sequences, thus expanding the reach of diagnostics into remote and/or economically disadvantaged areas.

Systems and methods for isothermal amplification of nucleic acids in a ratiometric manner are disclosed. Methods of isothermal amplification of nucleic acids are convenient for areas that lack reliable electricity and/or funding for precision lab equipment. However, these methods typically result in non-ratiometric amplification of target sequences, thus preventing quantitative analysis or application of the results, such as for diagnostic testing. Systems and methods herein provide a method of isothermal amplification that ratiometrically amplifies target sequences, thus expanding the reach of diagnostics into remote and/or economically disadvantaged areas.

The present invention provides, among other things, methods of quantitating mRNA capping efficiency, particularly for mRNA synthesized in vitro. In some embodiments, the methods comprise chromatographic methods of quantifying capping efficiency and methylation status of the caps.

The present invention provides, among other things, methods of quantitating mRNA capping efficiency, particularly for mRNA synthesized in vitro. In some embodiments, the methods comprise chromatographic methods of quantifying capping efficiency and methylation status of the caps.