Thermo Scientific NanoDrop One User manual

NanoDrop Micro-UV/Vis Spectrophotometers
NanoDrop One
User Guide
269-309102 Revision A May 2017

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For International Support, please contact:
Contact your local distributor. For contact
information go to:
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The contents of this document are subject to change without notice. All technical information in this
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WARNING Avoid an explosion or fire hazard. This instrument or accessory is not
designed for use in an explosive atmosphere.

Thermo Scientific NanoDrop One User Guide iii
C
Chapter 1 About the Spectrophotometer. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .1
Instrument Models and Features . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2
Optional Accessories . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5
Register Your Instrument . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6
Update Software . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7
Detection Limits for All Applications . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9
Measure dsDNA, ssDNA or RNA. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13
Measure dsDNA, ssDNA or RNA. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13
Nucleic Acid Reported Results . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 16
Setting for Nucleic Acid Measurements. . . . . . . . . . . . . . . . . . . . . . . . . . . . . 17
Calculations for Nucleic Acid Measurements. . . . . . . . . . . . . . . . . . . . . . . . . 18
Measure Microarray. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 23
Measure Microarray Samples. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 23
Microarray Reported Results . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 27
Settings for Microarray Measurements. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 28
Calculations for Microarray Measurements . . . . . . . . . . . . . . . . . . . . . . . . . . 32
Measure using a Custom Factor. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 35
Measure Nucleic Acid using a Custom Factor . . . . . . . . . . . . . . . . . . . . . . . . 35
Custom Factor Reported Results . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 37
Settings for Nucleic Acid Measurements using a Custom Factor . . . . . . . . . . 39
Detection Limits for Nucleic Acid Measurements using a Custom
Factor. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 39
Measure Oligo DNA or Oligo RNA . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 41
Measure Oligo DNA or Oligo RNA . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 41
Oligo Reported Results . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 45
Settings for Oligo DNA and Oligo RNA Measurements . . . . . . . . . . . . . . . . 47
Detection Limits for Oligo DNA and Oligo RNA Measurements . . . . . . . . . 48
Calculations for Oligo DNA and Oligo RNA Measurements . . . . . . . . . . . . 49
Measure Protein A280. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 53
Measure Protein Concentration at A280 . . . . . . . . . . . . . . . . . . . . . . . . . . . . 53
Protein A280 Reported Results . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 57
Settings for Protein A280 Measurements. . . . . . . . . . . . . . . . . . . . . . . . . . . . 59
Detection Limits for Protein A280 Measurements. . . . . . . . . . . . . . . . . . . . . 64
Calculations for Protein A280 Measurements . . . . . . . . . . . . . . . . . . . . . . . . 65
Contents

Contents
iv NanoDrop One User Guide Thermo Scientific
Measure Proteins and Labels . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 69
Measure Labeled Protein Samples . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 69
Proteins & Labels Reported Results. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 72
Settings for Proteins and Labels Measurements . . . . . . . . . . . . . . . . . . . . . . . 74
Detection Limits for Proteins and Labels Measurements . . . . . . . . . . . . . . . . 76
Calculations for Proteins and Labels Measurements. . . . . . . . . . . . . . . . . . . . 77
Measure Protein A205. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 79
Measure Protein Concentration at A205 . . . . . . . . . . . . . . . . . . . . . . . . . . . . 79
Protein A205 Reported Results . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 82
Settings for Protein A205 Measurements. . . . . . . . . . . . . . . . . . . . . . . . . . . . 83
Calculations for Protein A205 Measurements . . . . . . . . . . . . . . . . . . . . . . . . 85
Measure Protein BCA . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 87
Measure Total Protein Concentration . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 87
Protein BCA Reported Results . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 96
Settings for Protein BCA Measurements . . . . . . . . . . . . . . . . . . . . . . . . . . . 100
Measure Protein Bradford . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 101
Measure Total Protein Concentration . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 101
Protein Bradford Reported Results . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 106
Settings for Protein Bradford Measurements . . . . . . . . . . . . . . . . . . . . . . . . 109
Measure Protein Lowry . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 111
Measure Total Protein Concentration . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 111
Protein Lowry Reported Results . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 114
Settings for Protein Lowry Measurements . . . . . . . . . . . . . . . . . . . . . . . . . . 118
Measure Protein Pierce 660. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 119
Measure Total Protein Concentration . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 119
Protein Pierce 660 Reported Results . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 124
Settings for Protein Pierce 660 Measurements . . . . . . . . . . . . . . . . . . . . . . . 127
Measure OD600 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 129
Measure OD600 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 129
OD600 Reported Results . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 133
Settings for OD600 Measurements . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 134
Calculations for OD600 Measurements . . . . . . . . . . . . . . . . . . . . . . . . . . . 137
Measure Custom . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 139
Measure using a Custom Method . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 139
Delete Custom Method. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 143
Custom Method Reported Results. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 144
Measure UV-Vis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 147
Measure UV-Vis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 147
UV-Vis Reported Results. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 150
Settings for UV-Vis Measurements . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 152

Contents
Thermo Scientific NanoDrop One User Guide v
Measure Kinetics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 155
Measure Kinetics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 155
Create Kinetics Method. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 158
Edit Kinetics Method . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 159
Kinetics Reported Results . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 161
Settings for Kinetic Measurements. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 166
Chapter 18 Learning Center . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .169
Micro-Volume Sampling—How it Works. . . . . . . . . . . . . . . . . . . . . . . . . . . . 170
Set Up the Instrument. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 172
Measure a Micro-Volume Sample . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 188
Measure a Sample Using a Cuvette . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 195
Prepare Samples and Blanks . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 199
Basic Instrument Operations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 205
NanoDrop One Home Screen. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 206
NanoDrop One Measurement Screens . . . . . . . . . . . . . . . . . . . . . . . . . . . . 210
NanoDrop One Data Viewer . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 221
NanoDrop One General Operations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 229
Instrument Settings . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 240
Acclaro Sample Intelligence. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 248
NanoDrop One Viewer Software . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 256
Viewer Home Screen. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 257
Manage Experiments and Associated Data. . . . . . . . . . . . . . . . . . . . . . . . . . 259
Manage Identifiers on a PC . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 269
Manage Custom Methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 274
Multimedia . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 286
Chapter 19 Maintenance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .289
Maintenance Schedule. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 290
Cleaning the Touchscreen . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 291
Maintaining the Pedestals . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 292
Cleaning the Pedestals . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 292
Reconditioning the Pedestals . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 294
Decontaminating the Instrument . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 296
Maintaining the Cuvette Sampling System . . . . . . . . . . . . . . . . . . . . . . . . . . . 299
Instrument Diagnostics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 300
Intensity Check . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 300
Performance Verification. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 302
Pedestal Image Check . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 307
Chapter 5 Safety and Operating Precautions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .301
Operating Precautions. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 302
Safety Information. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 303

Contents
vi NanoDrop One User Guide Thermo Scientific
Chapter 6 About this Help System . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .311
Chapter 7 Technical Support. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .313

Thermo Scientific NanoDrop One User Guide 1
1
About the Spectrophotometer
The Thermo Scientific™ NanoDrop™ One is a compact,
stand-alone UV-Visible spectrophotometer developed for
micro-volume analysis of purified nucleic acids and a wide
variety of proteins. The patented sample retention system
enables the measurement of highly concentrated samples
without the need for dilutions.
The NanoDrop One system comes with preloaded software and
a touchscreen display. The instrument can be connected to an
optional printer with a USB cable or to a remote printer
through an Ethernet connection or wireless network.
NOTICE Before operating a NanoDrop One instrument, please read the safety and
operating precautions and then follow their recommendations when using the instrument.
Instrument Models and Features
There are two models available for the
NanoDrop One spectrophotometer...
Register Your Instrument
Register your instrument to receive e-mail updates on
software and...
Optional Accessories
A number of accessories are available for the
NanoDrop One instruments...
Update Software
Quickly and easily download the latest
NanoDrop One software...

1About the Spectrophotometer
Instrument Models and Features
2NanoDrop One User Guide Thermo Scientific
Instrument Models and Features
There are two models
available for the
NanoDrop One
spectrophotometer—the
NanoDrop One and the
NanoDrop OneC. Both
models include the patented
micro-volume sample
retention system and general
features. The
NanoDrop OneCmodel also
features a cuvette holder for
analyzing dilute samples using
standard UV-visible cuvettes.
Both instruments come with a
built-in, 7-inch Android
high-resolution touchscreen
preloaded with easy-to-use
instrument control software.
The NanoDrop One software
is loaded with features to
integrate with and simplify
your daily workflows.
1Locate the instrument away from air vents and exhaust fans to minimize evaporation
NanoDrop One Spectrophotometer NanoDrop OneCSpectrophotometer
Arm
Pedestal
Cuvette
holder

1 About the Spectrophotometer
Instrument Models and Features
Thermo Scientific NanoDrop One User Guide 3
Touchscreen
1Two more USB-A ports are located on instrument back panel
Touchscreen can slide left or
right to accommodate personal
preference, and tilt forward or
back for optimal viewing
USB-A port1
Pedestals
Touchscreen

1About the Spectrophotometer
Instrument Models and Features
4NanoDrop One User Guide Thermo Scientific
NanoDrop One Software with Acclaro Sample Intelligence Technology
NanoDrop OneCModel Additional Features
The Thermo Scientific™ Acclaro™ Sample
Intelligence technology built into the
NanoDrop One instruments provides these
exclusive features to help you assess sample
integrity:
• contaminant analysis to help qualify a sample
before use in downstream applications
• on-demand technical support for
measurements that are atypical or very low
concentration
• invalid result alerts (a column sensor
monitors for the presence of bubbles or
reflective particles that can compromise
measurement results)
The NanoDrop OneCmodel
includes a cuvette holder for
measuring dilute samples,
colorimetric assays, cell
cultures and kinetic studies.
The cuvette system has these
additional features:
• extended lower detection
limits
• 37 °C heater option for
temperature-sensitive
samples and analyses
• micro-stirring option to
ensure sample
homogeneity and support
kinetic studies
For details, see Measure a
Sample using a Cuvette.
Instrument light path
Cuvette holder

1 About the Spectrophotometer
Optional Accessories
Thermo Scientific NanoDrop One User Guide 5
Optional Accessories
A number of accessories are available for the NanoDrop One instruments. To order an
accessory, contact your local distributor or visit our website.
DYMO™ LabelWriter™ 450 USB Label Printer
Prints two 5/16-in x 4-in self-adhesive labels for transferring sample data directly into
laboratory notebooks or posting on bulletin boards. The software allows printing of data from
each sample measurement or from a group of samples logged and measured together.
The printer connects to the instrument (front or back) via a USB cable (included).
PR-1 Pedestal Reconditioning Kit
PV-1 Performance Verification Solution
Liquid photometric standard used to check instrument performance. For more information,
see Performance Verification.
Specially formulated conditioning compound that can be
applied to the pedestals to restore them to a hydrophobic state
(required to achieve adequate surface tension for accurate
sample measurements). The kit includes conditioning
compound and applicators. For more information, see
Reconditioning the Pedestals.

1About the Spectrophotometer
Register Your Instrument
6NanoDrop One User Guide Thermo Scientific
Register Your Instrument
Register your instrument to receive e-mail updates on software and accessories
for the NanoDrop One instruments. An Internet connection is required for
registration.
To register your instrument
1. Do one of the following:
–FromtheNanoDrop One Viewer software running on a personal
computer (PC) that is connected to the Internet, open the Help menu
and choose NanoDrop One Website.
– From any PC that is connected to the Internet, use any web browser to
navigate to our website.
2. On the website, locate NanoDrop One Registration and follow the
instructions to register the instrument.

1 About the Spectrophotometer
Update Software
Thermo Scientific NanoDrop One User Guide 7
Update Software
Quickly and easily download and install the latest NanoDrop One software and
release notes from our website. Follow the steps to update or upgrade the software
on your local instrument and/or install or update the NanoDrop One Viewer
software on a personal computer (PC). An Internet connection is required to
download software.
To install or update NanoDrop One Viewer software
1. Do one of the following:
• To install the Viewer software on a computer for the first time, open any
web browser and find the NanoDrop website.
• To update or upgrade the Viewer software, from the Viewer Home screen,
open the Help menu and choose NanoDrop One Website to open our
website.
2. On the NanoDrop website, locate the software downloads page.
3. Select to download NanoDrop One (PC) Viewer software (English version)
and follow the instructions to download and run the installer. (A computer
restart is required after the installer completes.)
4. To add a language, including software and Help systems, download and run the
language pack installer (English must be installed first). (No computer restart is
required after a language installer completes.)

1About the Spectrophotometer
Update Software
8NanoDrop One User Guide Thermo Scientific
To update or upgrade NanoDrop One instrument software
1. Do one of the following:
–FromtheNanoDrop One Viewer software, open the Help menu and
choose NanoDrop One Website to open our website.
– From any personal computer that is connected to the Internet, navigate to
the NanoDrop website.
2. Insert a USB device such as a memory stick into a USB port on the computer.
3. On the NanoDrop website, locate the software downloads page, select to
update or upgrade NanoDrop One operating software (English version) and
follow the instructions to download the installer to the USB device.
4. To add a language, including software and Help systems, download the
language pack installer(s) to the USB device.
5. Insert the USB device into any USB port on the NanoDrop One instrument.
6. From the instrument Home screen, tap (Settings) > System > Update
Software.
If the USB device contains more than one version of the installer, a message is
displayed. Select the version to install (English installer must be run first) and
tap Update. (An instrument restart is required after the English installer
completes.)
When the installation is complete, a message similar to the following appears
next to the Update Software button:
Version: 1.2.0 (currently installed version of instrument operating software)
Database version: 1 (version of NanoDrop One database on this instrument)
7. To add a language, including software and Help systems, tap Update Software
again,select the language and version to install and tap Update. (No
instrument restart is required after a language installer completes.)
Note: To change the language, tap Language, select an installed language and tap
OK. (An instrument restart is required after you change the language.)

Thermo Scientific NanoDrop One User Guide 9
2
Detection Limits for All Applications
Detection limits for standard applications
Note Detection limits provided in the tables below are approximate and apply to
micro-volume measurements only; they are based on the instrument’s photometric
absorbance range (10 mm equivalent) of 0–550 A. For measurements with 10 mm
pathlength cuvettes, the photometric absorbance range is 0–1.5 A.
Sample Type Lower Detection Limit Upper Detection Limit Typical Reproducibilitya
dsDNA 2.0 ng/μL (pedestal)
0.20 ng/μL (cuvette)
27,500 ng/μL (pedestal)
75 ng/μL (cuvette)
±2.0 ng/μL for sample concentrations
between 2.0 and 100 ng/μL samples;
±2% for samples >100 ng/μL
ssDNA 1.3 ng/μL (pedestal)
0.13 ng/μL (cuvette)
18,150 ng/μL (pedestal)
49.5 ng/μL (cuvette)
±2.0 ng/μL for sample concentrations
between 2.0 and 100 ng/μL samples;
±2% for samples >100 ng/μL
RNA 1.6 ng/μL (pedestal)
0.16 ng/μL (cuvette)
22,000 ng/μL (pedestal)
60 ng/μL (cuvette)
±2.0 ng/μL for sample concentrations
between 2.0 and 100 ng/μL samples;
±2% for samples >100 ng/μL
DNA Microarray
(ssDNA)
1.3 ng/μL (pedestal)
0.13 ng/μL (cuvette)
495 ng/μL (pedestal)
49.5 ng/μL (cuvette)
±2.0 ng/μL for sample concentrations
between 2.0 and 100 ng/μL samples;
±2% for samples >100 ng/μL

2
Detection Limits for All Applications
10 NanoDrop One User Guide Thermo Scientific
Purified BSA by
Protein A280
IgG by Protein
A280
0.06 mg/mL (pedestal)
0.006 mg/mL (cuvette)
0.03 mg/mL (pedestal)
0.003 mg/mL (cuvette)
825 mg/mL (pedestal)
402 mg/mL (pedestal)
±0.10 mg/mL (for 0.10–10 mg/mL samples);
±2% for samples >10 mg/mL
Purified BSA by
Proteins & Labels
0.06 mg/mL (pedestal)
0.006 mg/mL (cuvette)
19 mg/mL (pedestal) ±0.10 mg/mL for 0.10–10 mg/mL samples
Protein BCA 0.2 mg/mL (20:1
reagent/sample volume)
0.01 mg/mL (1:1
reagent/sample volume)
8.0 mg/mL (pedestal)
0.20 mg/mL (cuvette)
2% over entire range
0.01 mg/mL over entire range
Protein Lowry 0.2 mg/mL (pedestal) 4.0 mg/mL (pedestal) 2% over entire range
Protein Bradford 100 μg/mL (50:1
reagent/sample volume)
15 μg/mL (1:1
reagent/sample volume)
8000 μg/mL
100 μg/μL
±25 μg/mL for 100–500 μg/mL samples
±5% for 500–8000 μg/mL samples
±4 μg/mL for 15–50 μg/mL samples
±5% for 50–125 μg/mL samples
Protein Pierce 660 50 μg/mL (15:1
reagent/sample volume)
25 μg/mL (7.5:1
reagent/sample volume)
2000 μg/mL
1000 μg/mL
±3 μg/mL for 50–125 μg/mL samples
±2% for samples > 125 μg/mL
±3 μg/mL for 25–125 μg/mL samples
±2% for samples >125 μg/mL
aBased on five replicates (SD=ng/μL; CV=%)
Sample Type Lower Detection Limit Upper Detection Limit Typical Reproducibilitya
Note To minimize instrument error with highly concentrated samples, make dilutions to
ensure that measurements are made within these absorbance limits:
• For micro-volume measurements, maximum absorbance at 260 nm (for nucleic acids)
or 280 nm (for proteins) should be less than 62.5 A.
• For measurements with 10 mm pathlength cuvettes, maximum absorbance at 260 nm
(or 280 nm for proteins) should be less than 1.5 A, which is approximately 75 ng/μL
dsDNA.

2
Detection Limits for All Applications
Thermo Scientific NanoDrop One User Guide 11
Detection limits for pre-defined dyes
Sample Type Lower Detection Limit Upper Detection Limita
aValues are approximate
Typical Reproducibilityb
bBased on five replicates (SD=ng/μL; CV=%)
Cy3, Cy3.5, Alexa Fluor
555, Alexa Fluor 660
0.2 pmol/μL (pedestal) 100 pmol/μL (pedestal) ±0.20 pmol/μL for sample
concentrations between 0.20 and 4.0
pmol/μL;
±2% for samples >4.0 pmol/μL
Cy5, Cy5.5, Alexa Fluor
647
0.12 pmol/μL (pedestal) 60 pmol/μL (pedestal) ±0.12 pmol/μL for sample concentrations
between 0.12 and 2.4 pmol/μL;
±2% for samples >2.4 pmol/μL
Alexa Fluor 488, Alexa
Fluor 594
0.4 pmol/μL (pedestal) 215 pmol/μL (pedestal) ±0.40 pmol/μL for sample concentrations
between 0.40 and 8.0 pmol/μL;
±2% for samples >8.0 pmol/μL
Alexa Fluor 546 0.3 pmol/μL (pedestal) 145 pmol/μL (pedestal) ±0.30 pmol/μL for sample concentrations
between 0.30 and 6.0 pmol/μL;
±2% for samples >6.0 pmol/μL

Thermo Scientific NanoDrop One User Guide 13
3
Measure dsDNA, ssDNA or RNA
Measure dsDNA, ssDNA or RNA
Use the dsDNA, ssDNA and RNA applications to quantify purified double-stranded (ds) or
single-stranded (ss) DNA or RNA samples. These applications report nucleic acid
concentration and two absorbance ratios (A260/A280 and A260/A230). A single-point
baseline correction can also be used.
To measure dsDNA, ssDNA or RNA samples
Measures the concentration of
purified dsDNA, ssDNA or RNA
samples that absorb at 260 nm.
Measure dsDNA, ssDNA or RNA
Reported Results
Settings
Detection Limits
Calculations
NOTICE
• Do not use a squirt or spray bottle on or near the instrument as liquids will flow into
the instrument and may cause permanent damage.
• Do not use hydrofluoric acid (HF) on the pedestals. Fluoride ions will permanently
damage the quartz fiber optic cables.

3
Measure dsDNA, ssDNA or RNA
14 NanoDrop One User Guide Thermo Scientific
Before you begin...
Before taking pedestal measurements with the NanoDrop One instrument, lift the instrument
arm and clean the upper and lower pedestals. At a minimum, wipe the pedestals with a new
laboratory wipe. For more information, see Cleaning the Pedestals.
To measure nucleic acid
1. From the Home screen, select the Nucleic Acids tab
and tap dsDNA, ssDNA or RNA, depending on the
samples to be measured.
2. Specify a baseline correction if desired.
3. Pipette 1–2 μL blanking solution onto the lower
pedestal and lower the arm, or insert the blanking
cuvette into the cuvette holder.
Tip: If using a cuvette, make sure to align the cuvette
light path with the instrument light path.
4. Tap Blank and wait for the measurement to
complete.
Tip: If Auto-Blank is On, the blank measurement
starts automatically after you lower the arm. (This
option is not available for cuvette measurements.)
5. Lift the arm and clean both pedestals with a new
laboratory wipe, or remove the blanking cuvette.
6. Pipette 1-2 μL sample solution onto the pedestal and
lower the arm, or insert the sample cuvette into the
cuvette holder.
7. Start the sample measurement:
–Pedestal:IfAuto-Measure is On, lower arm; if
Auto-Measure is off, lower arm and tap Measure.
– Cuvette: Tap Measure.
When the sample measurement is completed, the
spectrum and reported values are displayed (see the
next section).
8. When you are finished measuring samples, tap
End Experiment.
9. Lift the arm and clean both pedestals with a new
wipe, or remove the sample cuvette.
Typical nucleic acid spectrum
Comparison of nucleic acid spectra with
and without two common contaminants