Nucleic acid quantification (NAQ) is the process of determining the concentration of DNA or RNA in a sample. Accurate NAQ is crucial in various molecular biology applications, including PCR, sequencing, cloning and gene expression analysis.
Learn how selecting the right tools can save time and money by preventing experimental failures in downstream applications.
Download this infographic to discover:
- How to select the most suitable NAQ technology
- A range of spectrophotometers and fluorometers to meet all throughput needs
- The importance of performing NAQ in PCR and NGS workflows
do you process per week? How many samples do you quantify at a time? Things to consider when selecting a tool Why perform NAQ? Using the correct amount of nucleic acid saves time and money, preventing downstream experimental failures. Downstream applications include: Cloning, NGS, qPCR, Ligation, PCR, Transfection, Microarrays, Sanger sequencing In PCR techniques: • Too little NA can result in failure to amplify the target • Too much NA used can lead to non- specific amplification Note: MIQE guidelines require input NA quality and purity to be reported in publication. MIQE - Minimum information for publication of quantitative real-time PCR experiment In NGS: • Accurate quantification of NA prior to library prep helps obtain high-quality reading data • Insufficient NA will result in low sequencing efficiency • NA overabundance will result in poor quality data Technologies used in NAQ How to select the most suitable technology How solutions from Thermo Fisher Scientific could help you If absorbance - blank ≤ 9X instrument precision (see instrument specifications), you cannot rely on the measurement. In this case switching to fluorescence will improve your result. Absorbance – UV-Vis Provides concentration of nucleic acid and protein by measuring the amount of light that it absorbs. Fluorescence In a fluorometric assay, dyes bind selectively to DNA, RNA, or protein of interest. Absorbance – UV-Vis InvitrogenTM Qubit™ 4 Fluorometer InvitrogenTM QubitTM Flex Fluorometer Thermo ScientificTM VarioskanTM LUX Multimode Microplate Reader Thermo ScientificTM NanoDropTM One & OneC Spectrophotometers Thermo ScientificTM NanoDropTM Eight Spectrophotometer Thermo ScientificTM MultiskanTM SkyHigh Microplate Spectrophotometer Thermo ScientificTM VarioskanTM LUX Multimode Microplate Reader Only 1-2μ sample needed for accurate quantification of one sample at a time Quantifies 8 samples at a time for improved productivity Offers high throughput and sample flexibility using 96384-well plates, cuvettes, or the Thermo Scientific™ μDrop™ Plate Versatile for both photometric and fluorescence quantification and offers increased flexibility addressing diverse research needs in laboratories Sensitivity and specificity one sample at a time Quantifies 8 samples with high sensitivity, accuracy and precision Versatile for both photometric and fluorescence quantification and offers increased flexibility addressing diverse research needs in laboratories For Research Use Only. Not for use in diagnostic procedures. © 2022 Thermo Fisher Scientific Inc. All rights reserved. All trademarks are the property of Thermo Fisher Scientific and its subsidiaries unless otherwise specified. COL35124 1122 Find out more about solutions that measure up at thermofisher.com/naq Increase throughput Increase throughput A few samples Multiskan SkyHigh NanoDrop 384-well plate A µDrop plate A pedestal, cuvette, array of pedestals 10-150 samples More than 150 samples Purified DNA/RNA Direct absorbance at 260 nm 96-well plate Multiskan SkyHigh Varioskan LUX Multiskan SkyHigh with µDrop plate NAQ determines the concentration of DNA or RNA in a sample, enabling the calculation of the correct amount of nucleic acid required for downstream applications. Sample Preparation DNA/RNA Purification DNA/RNA Quantification Downstream Application e.g. PCR Data Analysis What is nucleic acid quantification (NAQ)? Did you know? DNA or RNA absorbance spectrum Fluorescence Absorbance Fluorescence* T
