RNA/DNA Quantification | Thermo Fisher Scientific

RNA and DNA quantification instruments offered by Thermo Fisher Scientific: Qubit, NanoDrop and plate reader platforms

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UV-Vis Spectrophotometers
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Detection technologies used for RNA/DNA quantification

DNA and RNA quantification, generally referred to as nucleic acid quantification, is commonly performed to determine the average concentration of DNA or RNA in a sample prior to downstream experiments. Sample purity is also an important consideration to accurately calculate the amount of DNA or RNA in a sample. There are two optical technologies commonly used to quantify nucleic acids: UV-Vis measurement and fluorescence measurement. Choosing the right technology for your samples and workflow results in accurate RNA or DNA quantification and can save significant time and money by helping to prevent downstream experimental failures.

	Photometry (UV-Vis)	Fluorescence
How is the optical signal generated?	The photometric measurement of nucleic acids is based on the intrinsic absorptivity properties of nucleic acids (DNA and RNA). When an absorption spectrum is measured, nucleic acids absorb light with a characteristic peak at 260 nm.	The fluorometric measurement of nucleic acids is based upon the use of fluorogenic dyes that bind selectively to DNA or RNA.
	Typical RNA/DNA absorbance spectrum.	Fluorescent dyes selectively bind to DNA, RNA or protein. Dyes only emit signal when bound to the target.
How is the optical signal measured?	The signal is measured by spectrophotometers or spectrometers. The attenuation in the light that reaches the detector after passing through the sample is measured in relation to the incident light and expressed as absorbance values of the sample in the solution.	The signal is measured by fluorometers. Sample is excited with filtered light (at the excitation wavelength, and the emitted light (at the emission wavelength) is recorded by a detector.

	Wavelength separation can take place before or after the light has passed the sample, and the optical light path can be horizontal or vertical.	Wavelength separation can take place in various ways (for example with filters or with monochromators)
How is the concentration of nucleic acids calculated?	Concentrations of nucleic acids can be directly calculated from their measured absorbance values at 260 nm, using the Beer-Lambert's equation: where: C= nucleic acid concentration in molar (M) A=UV absorbance in absorbance units (AU) ε=wavelength-dependent molar absorptivity coefficient (or extinction coefficient) in M^-1cm^-1 L= light path in cm (cm) This concentration calculation is automated in many instruments.	Concentrations of nucleic acids are measured using the fluorescence signal of the sample and a calibration curve is generated from standard samples of known concentration and fit to appropriate regression models. Typical Fluorescence Standard Curve. The limit of detection and linear response of the measurements are specific to each assay.
What are the advantages?	It is simple—no sample preparation, dyes, or standards are required Can provide direct measurements of purity ratios—A260/280 and A260/230 Can provide information on contaminants—can identify non-nucleic acid contamination in samples (proteins, phenol, guanidine salts) as well as mammalian DNA/RNA contamination and provide corrected concentrations (applicable to NanoDrop One/One^C/Eight instruments)	It is specific—performed measurement is selective for DNA or RNA It is sensitive—can measure pg/mL; it is the recommended method for very diluted nucleic acid samples It is accurate despite contamination being present in the sample, including nucleic acid contaminants
What are the disadvantages?	It is not selective—uses software algorithms to distinguish mammalian DNA and RNA It has limited sensitivity—detection limits are higher than fluorescence-based methods	It takes longer—reagent and sample preparation are required No purity information is provided

We offer a range of instruments to perform nucleic acid quantification using either photometry of fluorescence measurements.

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Choice instruments for photometric UV-Vis RNA/DNA quantification

Well-known Thermo Scientific instruments for UV-Vis RNA/DNA quantification include the NanoDrop One/One^C Spectrophotometer for convenient, single-sample microvolume analysis, the NanoDrop Eight Spectrophotometer for 8-sample microvolume analysis, and choice of the Multiskan SkyHigh Microplate Spectrophotometer or the Varioskan LUX Multimode Reader for up to 16 microvolume samples as well as 96-384-well plates. The table below provides additional detail on instrument performance.

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Choice instruments for fluorescence RNA/DNA quantification

High-performing and popular instruments for performing fluorescence RNA or DNA quantification include Invitrogen Qubit 4 Fluorometer for single-sample microvolume fluorescence measurements, Invitrogen Qubit Flex Fluorometer for medium-throughput fluorescence measurements and the Thermo Scientific Varioskan LUX Multimode Microplate Reader for 96-1536-well microplate monochromator-based fluorescence measurements. The table below provides additional detail on instrument performance.

	Qubit 4	Qubit Flex	Varioskan LUX

What is the lower concentration limit (dsDNA)?	0.005 ng/µL*	0.005 ng/µL*	Depends on the Quant-iT assay kit, standard concentrations and curve fittings used** See details0.005 ng/µL
What is the upper detection limit (dsDNA)?	4000 ng/µL*	4000 ng/µL*	Depends on the Quant-iT assay kit and assay conditions
What are the available sample formats?	Qubit assay tubes See detailsLow-fluorescence, thin-walled polypropylene tubes	Qubit Flex tube strips See detailsLow-fluorescence, thin-walled PCR tube strips	96-1536 well plates
How many samples can be measured at a time? (throughput)	1	Up to 8	Dependent on assay format See details up to 96 with 96-well plates up to 384 with 384 well plates up to 1536 with 1536 well plates
What volume of sample is required to measure?	1-20 µL	1-20 µL	Dependent on assay format, fluorophore that is measured, among others
How much time does it take to measure 1 sample?	<3 seconds	~3 seconds for 8 samples	Around 1.3 minutes for 16 microvolume samples in the µDrop Plate See detailsWhen performed with the user interface touch screen or SkanIt software (fast mode)
Does it give information on sample purity or detect contaminants?	No	No	No
Does it distinguish between DNA and RNA in the sample?	Yes See detailsUsing fluorescent dyes that are specific for DNA or RNA	Yes See detailsUsing fluorescent dyes that are specific for DNA or RNA	Yes See detailsUsing fluorescent dyes that are specific for DNA or RNA
Does it provide a RNA quality and integrity score?	Yes, when used with Qubit RNA IQ Assay Kit	Yes, when used with Qubit RNA IQ Assay Kit	No
Does it support oligonucleotide quantification?	Yes See detailsWith ssDNA assay kit	Yes See detailsWith ssDNA assay kit	Yes See detailsWith Quant-iT OliGreen assay kit
Does it provide pre-configured protocols?	Yes See detailsThere are pre-configured protocols in the user interface touch screen	Yes See detailsThere are pre-configured protocols in the user interface touch screen	Yes See detailsThere are pre-configured protocols in the SkanIt Cloud Library, free with SkanIt Software
Does it connect to the Thermo Fisher Connect?	Yes	Yes	No
Does it provide 21CFR Part 11 compliance?	No	Yes, with optional software	Yes See detailswith SkanIt software Drug Discovery Edition
Does it offer robotic automation compatibility?	No	No	Yes
What is the overall ease of use?	++++	++++	++
What is the basic protocol?	See details Prepare working solutions of reagent using on-board Reagent Calculator Pipette samples and standards according to instructions shown on touch screen Results are available within seconds	See details Prepare working solutions of reagent using on-board Reagent Calculator Pipette samples and standards according to instructions shown on touch screen Results are available within seconds	See details Prepare reagents, sample(s) and standards according to specific Quant-iT kit instructions Pipette them in microplates Either select pre-built protocol from SkanIt Cloud Library or set-up assay using SkanIt Software Measure the plates with the instrument
What is the instrument price category?	Low	Low-Medium	High
What are the ownership costs?	Requires Qubit Assays Requires thin-walled polypropylene tubes See detailsRecommended: Qubit tubes	Requires Qubit Assays Requires PCR tube strips See detailsRecommended: Qubit Flex tube strips	Requires Quant-It assays Requires fluorescence-compatible plates See detailsRecommended: non-treated Nunc plates
* Reported here are the lower and upper quantitation ranges measured with Qubit dsDNA High sensitivity (HS) kit (Q32851) Qubit dsDNA Broad Range (BR) kit (Cat. No. Q32850) respectively and using variable sample volume (1-20 µL). ** Limit of Detection (LOD) using the Quant-iT dsDNA Assay HS Kit (Cat. No. Q33120) as instructed. LOD estimated according to IUPAC, based on the precision of the blanks (3*SD). Results will vary depending on the curve fittings, standard concentrations and used kits.

UV-Vis and Vis Spectrophotometry

Meet your analytical challenges with our complete line of ultraviolet-visible (UV-Vis) spectrophotometers. Our award-winning designs and user-friendly software help you quantify, assess purity, and more.

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Fluorometers and Fluorescence

Use fluorometers to quantify, detect and monitor analytes and their reactions by measuring the intensity of the fluorescent signal from dyes attached to biological molecules as well as naturally fluorescent molecules based on signature excitation (Ex) and emission (Em) wavelengths.

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Ordering information

UV-Vis spectrophotometers

Catalog #	Name	Price
ND-ONE-W	NanoDrop One Microvolume UV-Vis Spectrophotometer with Wi-Fi	Request A Quote
ND-ONEC-W	NanoDrop OneC Microvolume UV-Vis Spectrophotometer with Wi-Fi	Request A Quote
NDE-GL	NanoDrop Eight Spectrophotometer	Request A Quote
NDEIGHT-LAPTOP	NanoDrop Eight Spectrophotometer with laptop	Request A Quote
A51119700DPC	Multiskan SkyHigh with touchscreen, cuvette and μDrop Duo Plate	Request A Quote

FAQs

Q: What is the difference between UV and Fluorescence DNA quantification? Which DNA quantification technology should I choose?
A: UV and Fluorescence technologies work differently to quantify DNA. UV quantification relies on the intrinsic absorptivity of DNA and RNA molecules, while fluorescence quantification uses dyes that specifically bind to your molecule of choice. With UV technology, quantification isn't as sensitive, but it has a broader dynamic range and also gives data about sample purity--plus it is faster because there is no reagent prep. With fluorescence technology you get higher sensitivity and molecule-specific data, but it has lower dynamic range and reagent prep is required. The technology you need will depend on what features are important for your lab and experimentation.
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Q: Which is the best DNA quantification instrument? Which is the best RNA quantification instrument?
A: All of the DNA instruments offered by Thermo Fisher Scientific are of excellent quality. To find the right instrument to quantify DNA or RNA in your lab, explore the detailed comparison of UV-Vis spectrophotometers and fluorometers. Things like sensitivity, throughput and budget may be initial considerations when selecting a DNA quantification instrument. You may also consider whether you need target specificity, sample purity information, RNA quality information, or broad dynamic range.

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Resources

Support

Nucleic Acid Quantification Support Center
Find tips, troubleshooting help, and resources for your nucleic acid quantification workflow.

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	NanoDrop One/One^c	NanoDrop Eight	Multiskan SkyHigh	Varioskan LUX

What is the lower detection limit (dsDNA)?	Pedestal: 2.0 ng/µL Cuvette: 0.2 ng/µL	2.0 ng/µL	Depends on sample format and volume used (in the case of microplates)* See details Cuvette: 0.45 ng/µL µDrop Plate: 9 ng/µL Microplates: 96 well plates (normal) 70 µL (minimum) 2.1 ng/µL 140 µL (good) 1.1 ng/µL 370 µL (maximal) 0.4 ng/µL 96 well plates (half-area) 35 µL (minimum) 2.1 ng/µL 60 µL (good) 1.2 ng/µL 180 µL (maximal) 0.5 ng/µL 384 well plates 20 µL (minimum) 2.5 ng/µL 40 µL (good) 1.3 ng/µL 120 µL (maximal) 0.5 ng/µL	Depends on sample format and volume used (in the case of microplates)* See details Cuvette: 0.15 ng/µL µDrop Plate: 3 ng/µL Microplates: 96 well plates (normal) 70 µL (minimum) 0.7 ng/µL 140 µL (good) 0.4 ng/µL 370 µL (maximal) 0.5 ng/µL 96 well plates (half-area) 35 µL (minimum) 0.7 ng/µL 60 µL (good) 0.4 ng/µL 180 µL (maximal) 0.1 ng/µL 384 well plates 20 µL (minimum) 0.8 ng/µL 40 µL (good) 0.4 ng/µL 120 µL (maximal) 0.2 ng/µL
What is the upper detection limit (dsDNA)?	Pedestal: 27 500 ng/µL Cuvette: 75 ng/µL	10000 ng/µL	Depends on sample format and volume used (in the case of microplates) See details Cuvette: 125 ng/µL µDrop Plate: 2500 ng/µL Microplates: 96 well plates (normal) 70 µL (minimum) 590 ng/µL 140 µL (good) 310 ng/µL 370 µL (maximal) 120 ng/µL 96 well plates (half-area) 35 µL (minimum) 560 ng/µL 60 µL (good) 320 ng/µL 180 µL (maximal) 110 ng/µL 384 well plates 20 µL (minimum) 690 ng/µL 40 µL (good) 350 ng/µL 120 µL (maximal) 120 ng/µL	Depends on sample format and volume used (in the case of microplates) See details Cuvette: 200 ng/µL µDrop Plate: 4000 ng/µL Microplates: 96 well plates (normal) 70 µL (minimum) 950 ng/µL 140 µL (good) 500 ng/µL 370 µL (maximal) 190 ng/µL 96 well plates (half-area) 35 µL (minimum) 900 ng/µL 60 µL (good) 520 ng/µL 180 µL (maximal) 190 g/µL 384 well plates 20 µL (minimum) 1100 ng/µL 40 µL (good) 570 ng/µL 120 µL (maximal) 200 ng/µL
What are the available sample formats?	Drop (1 µL) cuvettes	Drop (1 µL)	96-384 well plates µDrop Plate (2-10 µL) cuvettes (not all models)	6-384 well plates µDrop Plate (2-10 µL) cuvettes (not all models)
How many samples can be measured at a time? (throughput)	One sample at a time	Up to 8	Dependent on assay format See details one sample at a time with cuvettes up to 16 with µDrop plate up to 96 with 96-well plates up to 384 with 384 well plates	Dependent on assay format See details one sample at a time with cuvettes up to 16 with µDrop plate up to 96 with 96-well plates up to 384 with 384 well plates
What volume of sample is required to measure?	1 µL	1 µL	As little as 2 µL depending on assay format See details 2-10 µL with µDrop Plate (smallest volume required) 70-300 µL with 96 well plates (normal) 20-100 µL with 384 well plates 1 mL with 10x10 mm cuvettes (70 µL for microcuvettes)	As little as 2 µL depending on assay format See details 2-10 µL with µDrop Plate (smallest volume required) 70-300 µL with 96 well plates (normal) 20-100 µL with 384 well plates 1 mL with 10x10 mm cuvettes (70 µL for microcuvettes)
How much time it takes to measure one sample?	8 seconds	< 20 seconds for 8 samples	Around 1 minute for 16 microvolume samples in the µDrop Plate See detailsWhen performed with the user interface touch screen or SkanIt software (fast mode)	Around 1.3 minutes for 16 microvolume samples in the µDrop Plate See detailsWhen performed with the user interface touch screen or SkanIt software (fast mode)
Does it provide spectral data of the sample?	Yes	Yes	Yes	Yes
Does it give information on sample purity?	Yes See detailsIt provides full spectral data (A260/A280 and A260/A230 ratios) as well as Acclaro Contaminant Analysis	Yes See detailsIt provides full spectral data (A260/A280 and A260/A230 ratios) as well as Acclaro Contaminant Analysis	Yes See detailsIt provides full spectral data (A260/A280 and A260/A230 ratios)	Yes See detailsIt provides full spectral data (A260/A280 and A260/A230 ratios)
Does it detect specific contaminants in the sample?	Yes See detailsAcclaro Sample Intelligence technology detects protein, phenol and guanidine salts and it gives true concentration of the nucleic acids	Yes See detailsAcclaro Sample Intelligence technology detects protein, phenol and guanidine salts and it gives true concentration of the nucleic acids	No See detailsSome information is available, based on the purity ratios	No See detailsSome information is available, based on the purity ratios
Does it distinguish between DNA and RNA in the sample?	Yes See detailsDistinguishes between mammalian DNA and mammalian RNA	Yes See detailsDistinguishes between mammalian DNA and mammalian RNA	No	No
Does it support oligonucleotide quantification?	Yes See detailsWith embedded applications and custom methods	Yes See detailsWith embedded applications and custom methods	Yes	Yes
Does it provide pre-configured protocols?	Yes See detailsThere are user interface touch screen and custom-protocols	Yes See detailsThere are user interface touch screen and custom-protocols	Yes See detailsThere are pre-configured protocols in the interface touch screen and SkanIt software Cloud Library, free with SkanIt Software	Yes See detailsThere are pre-configured protocols in the SkanIt software Cloud Library, free with SkanIt Software
Does it connect to the Thermo Fisher Connect?	Yes	No	Yes	No
Does it provide 21CFR Part 11 compliance?	Yes, with optional software	Yes, with optional software	Yes See detailswith SkanIt software Drug Discovery Edition	Yes See detailswith SkanIt software Drug Discovery Edition
Does it offer robotic automation compatibility?	No	No	Yes	Yes
How is the instrument operated?	Via a touch screen user interface or via the NanoDrop One PC control software	With a PC software (Nanodrop Eight Operating software)	Via a touch screen user interface or the PC software (SkanIt software)	Via a PC software (SkanIt software)
What is the overall ease of use?	++++	++++	++++	+++
What is the basic protocol?	See details Pipette buffer on pedestal and blank instrument Pipette sample directly on pedestal Follow instructions on touchscreen to run protocol Results are available within seconds	See details Pipette buffer on all 8 pedestals and blank instrument Pipette samples directly on 8 pedestals Run protocol using PC software Results are available within seconds	See details Pipette blank(s) and samples in microplates or µDrop plate Choose how you want to run your assay: Choose the pre-configured assay using the touch screen Choose the pre-built assay in the SkanIt Cloud Library within SkanIt Software Set up the assay using SkanIt Software Results are available quickly	See details Pipette blank(s) and samples into microplates or µDrop plate Either choose the pre-built protocol in the SkanIt Cloud Library or set-up the assay with SkanIt Software Results are available quickly
What is the instrument price category?	Medium	Medium	Medium	High
What are the ownership costs?	No recurring costs	No recurring costs	If using µDrop Plate, no recurring costs; otherwise UV-compatible clear plates	If using µDrop Plate, no recurring costs; otherwise UV-compatible clear plates
* Theoretical limit of detection (LOD), estimated according to IUPAC, based on the precision of the blanks (3*SD), from the precision specifications of Multiskan SkyHigh and Varioskan LUX microplate readers.