ABI Prism® 7900HT Sequence Detection System ... - OpenWetWare

Example Results The following figure illustrates a typical dissociation curve from an experiment run to
detect non-specific amplification in cDNA samples.
Before You Begin
Using SDS
Online Help
Examples in This
Chapter
-R n
-6.0
-5.0
-4.0
-3.0
-2.0
-1.0
Primer Dimer
T m =74.9°C
Main Product
T m = 80.5 °C
0.0
60 65 70 75 80
Temperature (°C)
85 90 95
The plot above displays the dual amplification peaks typical of primer-dimer formation.
The amplification from the specific product is displayed with a T m of 80.5 °C, while the
primer-dimer product has a characteristically lower T m of 74.9 °C.
For specific instructions on any procedure described within this section, refer to the
online help accompanying the SDS software. To get help at any time during the
procedure, click a help button ( ) located within the dialog box or window in which
you are working.
The illustrations and screenshots that appear within this chapter were created from a
plate run to determine the purity of a β-actin amplification in unknown samples. Each
well of the plate contains SYBR Green 1 dye, forward and reverse primers, and
genomic DNA known to contain complimentary binding sites.
Real-Time Analysis 6-19