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At a wavelength of 260 nm, the average extinction coefficient for double-stranded DNA is 0.020 (μg/mL) −1 cm −1, for single-stranded DNA it is 0.027 (μg/mL) −1 cm −1, for single-stranded RNA it is 0.025 (μg/mL) −1 cm −1 and for short single-stranded oligonucleotides it is dependent on the length and base composition.
Upon binding to DNA, the dye molecules assume a more rigid shape and increase in fluorescence by several orders of magnitude, most likely due to intercalation between the bases. [ 9 ] [ 10 ] The Qubit fluorometer, a device designed to measure fluorescence signals from samples, operates by correlating these signals with known concentrations of ...
DNA sequencing; Expression cloning; Fluorescence in situ hybridization; Lab-on-a-chip; Comparison of nucleic acid simulation software; Northern blot; Nuclear run-on assay; Radioactivity in the life sciences; Southern blot; Differential centrifugation (sucrose gradient) Toeprinting assay; Several bioinformatics methods, as seen in list of RNA ...
Quantitative PCR can also be applied to the detection and quantification of DNA in samples to determine the presence and abundance of a particular DNA sequence in these samples. [3] This measurement is made after each amplification cycle, and this is the reason why this method is called real time PCR (that is, immediate or simultaneous PCR).
Gene expression and RNA quantification studies have benefited from the increased precision and absolute quantification of dPCR. [91] RNA quantification can be accomplished via RT-PCR, wherein RNA is reverse-transcribed into cDNA in the partitioned reaction itself, and the number of RNA molecules originating from each transcript (or allelic ...
The other major high throughput technique employed in the quantification of specific RNA molecules is quantitative PCR, after reverse transcription of the RNA to cDNA. Several different short single-stranded DNA molecules (oligonucleotides) are used in a branched DNA-assay. The capture and capture-extender oligonucleotide bind to the target ...
The assay is using S1 nuclease, which degrades single-stranded DNA and RNA into oligo- or mononucleotides, leaving intact double-stranded DNA and RNA. In the nuclease hybridization assay, the oligonucleotide analyte is captured onto the solid support such as a 96-well plate via a fully complementary cutting probe.
RNA-Skim RNA-Skim: a rapid method for RNA-Seq quantification at transcript-level. rSeq rSeq is a set of tools for RNA-Seq data analysis. It consists of programs that deal with many aspects of RNA-Seq data analysis, such as read quality assessment, reference sequence generation, sequence mapping, gene and isoform expressions (RPKMs) estimation, etc.
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