3.
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 3.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 3.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 3.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.
Materials to Be Supplied By User
•
thin-walled PCR tubes (Cat.# E4942 or Axygen Cat.# PCR-05-C, available from
Fisher or VWR)
3.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.
Blank and Standard Sample Preparation
1. Prepare 1X TE buffer by diluting 20X TE Buffer (pH 7.5) to 1X with
nuclease-free water. For example, combine 200µl of 20X TE Buffer and
3,800µl of nuclease-free water to prepare 4ml of 1X TE buffer.
Promega Corporation
·
2800 Woods Hollow Road
·
Madison, WI 53711-5399 USA
Toll Free in USA 800-356-9526
·
Phone 608-274-4330
·
Fax 608-277-2516
·
www.promega.com
Part# TM396
Printed in USA.
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