Nucleic Acid (DNA/RNA) Detection, Quantitation, and Analysis

Quantitate and analyze nucleic acids in a microplate format

What are nucleic acids?

Nucleic acids are large, complex biomolecules found in cells. They serve as the genetic material of living organisms and contain the instructions for the formation and function of all living things. They are generally found in two naturally occurring forms: DNA (deoxyribonucleic acid) and RNA (ribonucleic acid).

DNA and RNA

DNA and RNA are composed of nucleotides, each of which consists of a 5-carbon sugar, a nitrogenous base, and a phosphate group. DNA is formed by a double strand of paired nucleotides, while ribonucleic acid (RNA) is typically a single strand. In DNA, the nucleotides are adenine, cytosine, guanine, and thymine, while RNA contains uracil instead of thymine. Sequences of DNA nucleotides are organized into units called genes, which contain the information encoding individual proteins. DNA for a gene is transcribed to make an RNA copy of the gene, which then serves as the template for synthesis of the protein.

DNA damage and mutations that alter the sequence of genes can result in malfunctioning proteins that disrupt normal cellular function. Cancer is a prime example of how genetic mutations can disrupt the normal regulation of cellular behavior, leading to uncontrolled cell growth. Research into the function of genes and the genetic changes that give rise to disease will continue to lead to new therapeutics.

Nucleic acid detection and quantification methods

Nucleic acid is often purified from cells as part of an ever-growing array of molecular biology methods, including sequencing and gene editing. Before they are used in downstream applications, nucleic acids are detected and quantitated using UV or fluorescence spectrophotometry. Traditionally measured individually in cuvettes, sample analysis is now routinely performed in microplates.

Molecular Devices provides a complete workflow solution for nucleic acid detection, quantitation, and analysis. Our application notes demonstrate the quantitation and analysis of nucleic acids in a microplate format, offering higher throughput compared to other methods, as well as automated calculation of results.

  • DNA/RNA absorbance measurements

    DNA/RNA Quantitation

    The absorbance of a DNA sample measured at 260 nm on a spectrophotometer or microplate reader can be used to calculate its concentration. Absorbance quantitation of DNA works on samples ranging from about 0.25 μg/mL to about 125 μg/mL in a microplate format.

    Learn how absorbance is measured on our absorbance microplate readers with our featured application notes:

    eBook: Nucleic acid & protein quantitation

    Streamline absorbance assays for nucleic acid & protein quantitation

    Streamline absorbance assays for nucleic acid & protein quantitation

    Absorbance microplate readers are widely used in basic research, drug discovery, bioassay validation, quality control, and manufacturing processes in pharmaceutical, biotech, food and beverage, and academic industries. These readers provide rapid and sensitive measurements of a variety of analytes across a wide range of concentrations for a wide range of assays, including ELISAs, microbial growth, detection of key compounds and contaminants, and protein quantitation.

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  • Fluorometric quantitation of nucleic acids

    Nucleic acid quantitation

    Quantitation of DNA is a critical step in molecular biology requiring accuracy, reliability, and the use of increasingly smaller sample volumes for applications such as next-generation sequencing. Compared to spectrophotometric DNA quantitation, the fluorometric method provides key advantages such as significantly increased sensitivity, high selectivity for double-stranded DNA (dsDNA) over single-stranded DNA (ssDNA) or RNA, and improved contaminant tolerance (protein and carbohydrate molecules).

    Here are a few application notes on fluorometric quantitation of nucleic acids that you may find of interest:

    SNP genotyping

    SNP Genotyping

    Genotyping is a process for analyzing genetic differences among individuals by examining their DNA sequences. Single nucleotide polymorphisms (SNPs) are one of the most common types of genetic variation, consisting of a single nucleotide mutation at a specific locus. SNP genotyping has proven very useful in identifying disease-related mutations in various species, and as a result many techniques for SNP detection have been developed.

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