Determining Optimum Dye Concentration - Applied Biosystems 7300 Installation And Maintenance Manual

Appendix C

Determining Optimum Dye Concentration

Determining Optimum Dye Concentration
Wear powder-free
IMPORTANT!
gloves while creating a custom pure
dye plate.
In the center wells of a 96-well plate, prepare a
1.
dilution series of the custom dye (for example,
25, 50, 100, 200, 400, 800, 1600, and 3200 nM
concentrations) using 50-µL volumes for the
7300/7500 system or 20-µL volumes for the
7500 Fast system.
Seal the wells of the reaction plate using an
2.
optical adhesive cover.
In the SDS software, click
3.
select File New).
In the New Document dialog box, click
4.
.
It is not necessary to configure detector,
Note:
sample, and method information for the plate
document. The purpose of the run is to establish
the correct working concentration for the dye by
viewing the intensity of the raw spectra produced
by the wells in the dilution series.
Load the prepared plate:
5.
Press the tray door to open it.
a.
Load the pure dye plate into the plate
b.
holder.
Press the tray door to move the drawer into
c.
the instrument.
Notes
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Applied Biosystems 7300/7500/7500 Fast Real-Time PCR System Installation and Maintenance Guide
(or
25 50 100 200
400 800 1600 3200