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.
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DNA/RNA absorbance measurements
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:
DNA/RNA absorbance measurements
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
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.
eBook: 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
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:
Fluorometric quantitation of nucleic acids
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
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.
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.
Resources of Nucleic acid (DNA/RNA) Quantitation and Analysis
Application Note
Sensitive RNA fluorescent quantitation with the Quant-iT RiboGreen RNA assay kit
Sensitive RNA fluorescent quantitation with the Quant-iT RiboGreen RNA assay kit
Accurate quantification of nucleic acid concentration is important for downstream applications including transfection, cloning, PCR, and next generation sequencing (NGS). Often, these…
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Application Note
Spectral Fusion™ Illumination technology for an extended dynamic range on the SpectraMax i3x Multi-Mode Microplate Reader
Spectral Fusion™ Illumination technology for an extended dynamic range on the SpectraMax i3x Multi-Mode Microplate Reader
Many microplate readers and spectrophotometers use a photomultiplier tube (PMT) to detect fluorescence and luminescence signals. PMTs work by initially detecting photons from the sample…
eBook
Virology and Vaccine Research Microplate Reader Solutions
Virology and Vaccine Research Microplate Reader Solutions
We understand what a challenging time this is for those who are working to get accelerated testing and medicines to the market as soon as possible and are committed to supporting scientists…
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eBook
The Ultimate Guide to Microplate Reader Solutions
The Ultimate Guide to Microplate Reader Solutions
Evaluating microplate readers doesn’t have to be overwhelming. First, consider your application needs. If your budget is modest, a single-mode reader dedicated to your main application may b…
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Application Note
Sensitive fluorescent quantitation of DNA with the Quant-iT PicoGreen dsDNA Assay Kit
Sensitive fluorescent quantitation of DNA with the Quant-iT PicoGreen dsDNA Assay Kit
Double-stranded DNA is typically quantitated in microplate readers by measuring the absorbance of the DNA solution at 260 nm. However, this method is only able to measure down to about 250…
Publications
Multiple Steps from Microplate-Reader Workstations
Multiple Steps from Microplate-Reader Workstations
Scientists take many roads to overcoming bottlenecks in workflows. For example, a workstation including a microplate reader can handle multiple steps, which improves efficiency and adds spee…
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eBook
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 bevera…
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Application Note
Low-volume, high-throughput DNA and protein detection on the SpectraMax ABS Plus Microplate Reader
Low-volume, high-throughput DNA and protein detection on the SpectraMax ABS Plus Microplate Reader
Nucleic acid and protein quantitation are essential measurements upstream of many sophisticated assays in genetics and molecular biology. Various methods have been developed to quantitate…
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Application Note
DNA and RNA absorbance measurements using SpectraMax Microplate Readers
DNA and RNA absorbance measurements using SpectraMax Microplate Readers
Ultraviolet (UV) measurements in microplates became possible when Molecular Devices introduced the first UV-capable microplate reader. Since then, microplate measurements of DNA, RNA, and…
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Application Note
Quantitation of dsDNA samples using the SpectraMax Quant dsDNA Assay Kits
Quantitation of dsDNA samples using the SpectraMax Quant dsDNA Assay Kits
As different types of test samples may contain different concentrations of DNA, selection of an appropriate fluorescent reagent with the required sensitivity and detection range is…
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Application Note
Detect SNPs with KASP genotyping technology on SpectraMax microplate readers
Detect SNPs with KASP genotyping technology on SpectraMax microplate readers
Genotyping is a process for analyzing genetic differences among individuals by examining their DNA sequences. The many methods available for genotyping enable researchers to investigate…
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