Quantus Fluorometer Operating Manual TM396
Quantus Fluorometer Operating Manual TM396
Quantus Fluorometer Operating Manual TM396
Quantus™ Fluorometer
Instructions for Use of Product
E6150
Revised 10/22
TM396
Quantus™ Fluorometer
All technical literature is available at: www.promega.com/protocols/
Visit the web site to verify that you are using the most current version of this Operating Manual.
E-mail Promega Technical Services if you have questions on use of this system: [email protected]
1. Description.............................................................................................................................................2
2. Product Components................................................................................................................................2
3. Specifications.........................................................................................................................................3
3.A. Precautions and Special Instructions.................................................................................................4
3.B. Environmental Requirements............................................................................................................4
7. Troubleshooting.................................................................................................................................... 12
7.A. Common Problems........................................................................................................................ 12
7.B. Error Messages ............................................................................................................................ 13
8. Appendix.............................................................................................................................................. 14
8.A. Updating Firmware........................................................................................................................ 14
8.B. Warranty and Service..................................................................................................................... 14
8.C. Certificate of Decontamination........................................................................................................ 15
8.D. Related Products.......................................................................................................................... 16
9. Summary of Changes............................................................................................................................. 16
Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 1
www.promega.com TM396 · Revised 10/22
1. Description
The Quantus™ Fluorometer is a dual-channel fluorometer for your personal quantitation workflow. Designed to provide
highly sensitive fluorescent detection when quantifying nucleic acids and proteins, the compact instrument is simple to
operate. The Quantus™ Fluorometer is optimized with preprogrammed settings for the QuantiFluor® Dye Systems
(QuantiFluor® dsDNA, RNA and ssDNA Systems) for nucleic acid quantitation and allows users the flexibility to create
their own methods and quantitation settings for other dyes within the proper excitation and emission wavelengths.
The Quantus™ Fluorometer is equipped with two fluorescence channels for nucleic acid and protein quantitation:
• Red fluorescence channel: Excitation 640nm shortpass (wavelengths up to 640nm), emission 660–720nm;
• Blue fluorescence channel: Excitation 495nm shortpass (wavelengths up to 495nm), emission 510–580nm.
2. Product Components
P R O D U CT S I Z E C A T. #
• Quantus™ Fluorometer
• USB Power Supply
• USB Cord
• Outlet Adapter Kit (4 adapters supplied for international compatibility)
• Quick Reference Guide
• Product Registration Card
Shipping and Storage Conditions: Ship and store Quantus™ Fluorometer at +5°C to +40°C under noncondensing conditions,
at up to 75% humidity.
The Quantus™ Software, which can be used to transfer data to a computer, can be downloaded at:
www.promega.com/quantusfluorometersoftware/
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3. Specifications
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3.A. Precautions and Special Instructions
Special Instructions:
• Failure to operate, store and use the Quantus™ Fluorometer as directed will invalidate the warranty.
• Direct plug-in power supply acts as a disconnect. To disconnect power, pull power supply from the outlet.
• Changes or modifications to this unit not expressly approved by the party responsible for compliance could void the
user’s authority to operate the equipment.
• Do not use this device in proximity to sources of strong electromagnetic radiation (e.g., unshielded intentional RF
sources), because they may interfere with the proper operation.
• Do not use this instrument for anything other than its intended use.
• If the instrument is used in a manner other than that specified by Promega, the protection provided by the equipment
may be impaired.
• Do not disassemble the unit.
• The Quantus™ Fluorometer accepts 0.5ml PCR tubes.
• Do not force larger tubes into the tube holder.
• Use caution around solvents because they may corrode the plastic case of the Quantus™ Fluorometer.
• Do not expose the Quantus™ Fluorometer to temperatures outside the specified range since damage to the unit may
occur.
• Avoid bubbles in your sample because bubbles may negatively affect the fluorescence reading.
• Always disconnect the power before cleaning or performing routine maintenance.
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4. Quantus™ Fluorometer Inspection and Setup
Upon receiving your Quantus™ Fluorometer, inspect everything carefully and make sure all items are present and undamaged.
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5. Operating the Quantus™ Fluorometer
The Quantus™ Fluorometer uses five buttons for simple navigation [one center button to confirm your choice (“Go”), and
four surrounding buttons to navigate up, down, left or right through the menus].
The instrument is designed for use with Promega QuantiFluor® Dye Systems for fluorescence-based quantitation of
dsDNA, ssDNA and RNA. A protocol for each QuantiFluor® Dye is listed on the Protocol screen. Each of these protocols
contains optimized calibration curve settings based on the blank, standard, sample and dye preparation described in
Section 5.A. The instrument also may be used with other fluorescent dyes capable of excitation and emission in the
proper wavelength range. If you are using a dye other than one of the QuantiFluor® Dyes, a different dye dilution or a
different standard or unknown sample preparation method, follow the instructions in Section 5.B to create a user-defined
protocol.
The Quantus™ Fluorometer uses a single-point calibration process. The fluorometer must be calibrated prior to the first
use of a protocol. If a protocol that has never been calibrated is selected, the Calibration screen will appear. If a protocol
has been calibrated, the Home screen will appear, and you can proceed to Section 5.D. Each Quantus™ Fluorometer
protocol has calibration values associated with the specific assay reagent. Once the protocol of interest is calibrated, the
Quantus™ Fluorometer will save the calibration values for future use and will not require recalibration. However, we
recommend performing a calibration with each new lot of dye reagent to ensure the most accurate measurement.
5.A. Calibrating the Quantus™ Fluorometer for Use With the QuantiFluor® Dye Systems
Follow this protocol to calibrate the Quantus™ Fluorometer prior to quantitation using the QuantiFluor® Dye Systems. The
table below lists the dilution factors used to prepare the standard samples for the various QuantiFluor® Dye Systems
when using the standard provided with the system. If using a different DNA or RNA sample as a standard, prepare the
standard sample at the final standard calibration (ng/tube) indicated below.
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QuantiFluor® Dye System Dilution of QuantiFluor® Dye
ONE dsDNA (Cat.# E4871), single-concentration standard
no dilution necessary (dye is pre-diluted)
curve
dsDNA (Cat.# E2670), single-concentration standard
1:400
curve
RNA (Cat.# E3310), high-concentration standard curve 1:400
RNA (Cat.# E3310), low-concentration standard curve 1:2,000
3. Prepare the nucleic acid standard in a 0.5ml PCR tube. Use the volume of supplied standard and volume of 1X TE
buffer indicated in the table below to prepare the standard.
Final
Starting Volume of Standard
Standard Volume of Working Calibration
QuantiFluor® Dye System Concentration Standard Solution (per tube)
ONE dsDNA (Cat.# E4871) 400ng/μl 1μl 200μl 400ng
dsDNA (Cat.# E2670) 100ng/μl 2μl 200μl 200ng
RNA (Cat.# E3310),
high-concentration 100ng/μl 5μl 200μl 500ng
standard curve
RNA (Cat.# E3310), First dilute 1:100
low-concentration 100ng/μl in 1X TE Buffer, 200μl 10ng
standard curve then use 10μl
ssDNA (Cat.# E3190),
high-concentration 100ng/μl 4μl 200μl 400ng
standard curve
ssDNA (Cat.# E3190), First dilute 1:100
low-concentration 100ng/μl in 1X TE Buffer, 200μl 10ng
standard curve then use 10μl
4. Prepare the blank sample for the QuantiFluor® ONE dsDNA System by adding 200μl of QuantiFluor® ONE dsDNA Dye
to a 0.5ml PCR tube.
Prepare the blank sample for all other QuantiFluor® Systems by adding 200μl of QuantiFluor® Dye working solution
prepared in Step 2 to a 0.5ml PCR tube.
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5.A. Calibrating the Quantus™ Fluorometer for Use With the QuantiFluor® Dye Systems (continued)
5. Prepare the standard and samples. For the QuantiFluor® ONE dsDNA System, add 1μl of the standard to 200μl of
QuantiFluor® ONE dsDNA Dye.
For all other QuantiFluor® Systems, add standard prepared in Step 3 to 200μl of QuantiFluor® Dye working solution
prepared in Step 2.
6. Mix three times by pipetting slowly. When using aerosol-resistant pipette tips, do not allow the pipette tip filter to
get wet. Alternatively, vortex tubes at a high setting for 10 seconds.
7. Optional: Centrifuge tubes at 2,000 × g for 5–10 seconds to collect liquid at the bottom of the tube and remove any
bubbles present.
8. Incubate tubes at room temperature for 5 minutes, protected from light.
Calibration Protocol
1. Select the desired QuantiFluor® Dye assay from the Protocol screen on the instrument. If this is the first time the
protocol has been selected, the Calibration screen will automatically appear. Otherwise, after selecting the desired
protocol, navigate to the Calibration screen.
2. Place the blank sample into the tube holder, and close the lid. Select Read Blank, and the fluorescence in relative
fluorescence units (RFU) for the blank sample will be displayed on the screen.
3. Place the standard sample into the tube holder, and close the lid. Select Read Std, and the fluorescence in RFU for
the standard sample will be displayed on the screen.
Note: Once both the blank and standard samples have been measured, the standard-to-blank ratio is automatically
calculated as a quality check to ensure the QuantiFluor® Dye working solution and blank and standard samples
were prepared properly. If the standard-to-blank ratio is lower than the minimal accepted ratio, the screen will
display “Invalid”, and the instrument will not quantitate the unknown samples until the blank and standard samples
are prepared properly. The minimum standard-to-blank ratio is:
• 50 for high-concentration standard curve or single standard curve protocols
• 4 for low-concentration standard curve protocols
• 3 for user-defined protocols
These ratios have been predetermined as a quality check to help ensure a successful measurement.
4. Save the calibration data by selecting Save. Proceed to Section 5.D.
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3. Enter the high-concentration standard value by choosing 1–1,000. Select the concentration units and sample volume.
Note: The standard value is entered when setting up the user-defined protocol. After calibration, you can change
the units and volume from the Main screen. Several factors will influence the standard value entered, including the
concentration units, mass of nucleic acid per tube and sample volume. The instrument firmware assumes a 200μl
final volume. For example, with the concentration units set to ng/μl, the sample volume set to 1μl, and the mass of
nucleic acid at 100ng per tube, the standard value entered is 100/200 = 0.5.
4. Choose the appropriate fluorescence channel:
Red: Excitation 640nm shortpass, emission 660–720nm
Blue: Excitation 495nm shortpass, emission 510–580nm
5. Select Save.
Note: To edit or delete a user-defined protocol, select Edit next to the protocol name.
5.C. Calibrating the Quantus™ Fluorometer for Use With Other Dye Chemistries
User-defined protocols must be calibrated using blank and standard samples prepared similarly to those described in
Section 5.A. You may choose another standard, but the instrument assumes the total assay volume in the tube is 200μl. If
less than 200μl is used, the sample may not be completely within the light path, causing inaccurate sample readings, and
the concentration calculation must be corrected, since the instrument calculates based on a 200μl assay volume.
1. Prepare the blank and standard samples as directed in Section 5.A. Alternatively, you may use a different standard
value in the user-defined protocol. We recommend a minimum assay volume of 200μl. Note that the instrument
software assumes a linear relationship when calculating results from a single-point setup (blank and standard). Be
sure to verify, through a dilution series, that any dye dilution or standard combination is accurate.
2. Place the blank sample into the tube holder, and close the lid. Select Read Blank, and the fluorescence in relative
fluorescence units (RFU) for the blank sample will be displayed on the screen.
3. Place the standard sample into the tube holder, and close the lid. Select Read Std, and the fluorescence RFU for the
standard sample will be displayed on the screen.
Note: Once both the blank and standard samples are measured, the standard-to-blank ratio is automatically
calculated. If the standard-to-blank ratio is less than 3, the screen will display “Invalid”, and the instrument will not
quantitate unknown samples until the blank and standard samples are prepared properly (see Section 5.A). User-
defined protocols use a minimum standard-to-blank ratio of 3 as the quality check.
4. Save the calibration data by selecting Save. Proceed to Section 5.D.
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5.D. Quantitating Samples
Note: If quantitating samples of higher concentration than the standard, dilute the sample to ensure it is within the linear
range of the standard.
1. Press any button to wake up the Quantus™ Fluorometer. The Home screen will appear, displaying the last protocol
used, the last concentration measurement and a list of menu items for navigation.
2. Navigate to the right side of the screen, and select Protocol from the menu list on the Home Screen.
3. Select the desired protocol.
4. Navigate to and select the Volume/Units button to enter the volume of sample (i.e., template volume) added to the
assay.
5. To enter the sample volume, press the up or down buttons to the desired volume. Choose from 1–10, 15, 20, 25, 50,
100, 150 or 200μl.
Note: This volume is the amount of sample that was added to the assay.
6. To select the concentration units used to display the data, press the up or down button. Choose ng/μl, ng/ml, μg/ml,
mg/ml or Auto.
Note: If Auto is selected, the instrument will automatically assign units that place the concentration value in the
1.0 to 1,000.0 range.
7. When the desired units are highlighted, press Enter.
8. When using the QuantiFluor® ONE dsDNA System, mix 1–20μl of unknown sample and 200μl of QuantiFluor® ONE
dsDNA Dye in a 0.5ml PCR tube. Mix well by pipetting or vortexing. Incubate reactions for 5 minutes at room
temperature, protected from light.
When using all other QuantiFluor® Systems, mix 1–20μl of unknown sample with 200μl of QuantiFluor® Dye working
solution in a 0.5ml PCR tube. Mix well by pipetting or vortexing.
9. Place the unknown sample into the tube holder, and close the lid. The instrument will automatically measure
fluorescence when the lid is closed, and the calculated nucleic acid concentration will be displayed.
5.E. Tools
The instrument is set to automatically measure fluorescence of the sample when the lid is closed. This streamlines the
workflow. However, this feature may be turned off in the Tools screen.
To display only raw RFU values in either the Blue or Red channels, select the Raw Measurement option in the Tools screen.
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5.F. Results History
The Quantus™ Fluorometer will save the last 50 measurements. These measurements are displayed on the History screen.
Page up or down using the menu list as needed to view the available history.
To erase the result history, select Clear from the menu list.
Results can be transferred to a connected computer as the measurements are read using the Quantus™ Software (see
Section 4.B). The default fields transferred include sample ID, protocol name, sample concentration data with units and
instrument status (e.g., OK, SAT or LOW). Additional fields also may be transferred by selecting these fields under the
Tools icon (top right corner of the software). These additional fields include transfer time (showing the date and time
when data were transferred to the software), raw measurement data, blank and standard calibration values, and sample
volume.
Alternatively, transfer the last 50 measurements from the instrument to the computer by selecting Transfer from the
History screen on the instrument.
Note: the Quantus™ Instrument must be connected to the computer to transfer data. Otherwise, a connection error will be
displayed in the software.
To export results from the Quantus™ Software, select Save/Save As at the bottom of the screen. The data will export as a
comma separated value (CSV) file, which can be opened for data manipulation and printing.
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7. Troubleshooting
For questions not addressed here, please contact your local Promega Branch Office or Distributor. Contact information
available at: www.promega.com. E-mail: [email protected]
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TM396 · Revised 10/22 www.promega.com
Symptoms Causes and Comments
Quantus™ Fluorometer powered up, but screen Unplug USB Cord from the instrument, and plug the cord back in,
was blank, could not be deciphered or did not making sure no buttons are pressed during this time.
respond when buttons were pressed
Quantus™ Fluorometer was plugged in, Instrument entered sleep mode. Press any button to wake up
but the screen was dark the instrument.
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7.B. Error Messages (continued)
8. Appendix
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TM396 · Revised 10/22 www.promega.com
8.C. Certificate of Decontamination
Disinfection and decontamination are required prior to shipping the instrument and instrument accessories for repair.
Returned instruments must be accompanied by a signed and dated Certificate of Decontamination attached to the outside
packaging of the instrument.
To disinfect and decontaminate: Unplug the instrument, and then wipe the outside surfaces with a cloth dampened with a
mild detergent. Do not submerge the instrument in water.
Failure to confirm disinfection and decontamination will result in decontamination charges before the instrument is
serviced.
Select (A) or (B):
A. I confirm that the returned items have not been contaminated by body fluids or toxic, carcinogenic, radioactive or
other hazardous materials.
B. I confirm that the returned items have been decontaminated and can be handled without exposing personnel to
health hazards.
Circle the type of material used in the instrument: Chemical Biological Radioactive**
Briefly describe the decontamination procedure performed:
Date:___________________________
Return Authorization Number:_____________________________________________________
Institution:__________________________________________________________________
Signature:___________________________________________________________________
Name (block capital letters):_______________________________________________________
** The signature of a Radiation Safety Officer is also required if the instrument was used with radioactive materials.
This instrument is certified by the undersigned to be free of radioactive contamination.
Date:________________________
Institution:__________________________________________________________________
Signature:___________________________________________________________________
Name (block capital letters): _______________________________________________________
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www.promega.com TM396 · Revised 10/22
8.D. Related Products
9. Summary of Changes
The following changes were made to the 10/22 revision of this document:
1. Edited text for Step 5, Section 5.A.
2. Revised the cover page, including moving Figure 1 to cover.
3. Updated document font and title.
4. Made minor text edits.
Products may be covered by pending or issued patents or may have certain limitations. Please visit our Web site for more information.
All prices and specifications are subject to change without prior notice.
Product claims are subject to change. Please contact Promega Technical Services or access the Promega online catalog for the most up-to-date information on
Promega products.
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TM396 · Revised 10/22 www.promega.com