Power SYBR Green PCR Master Mix and RT-PCR Protocol (PN ...

Chapter 1
Introduction
Using
Dissociation
Curves
General Information
The following systems enable you to run dissociation curves to detect
nonspecific amplification through computer software:
• Applied Biosystems 7900HT Real-Time PCR System
• Applied Biosystems 7500 Real-Time PCR System
• Applied Biosystems 7300 Real-Time PCR System
• ABI PRISM ® 7700 SDS
• ABI PRISM ® 7000 SDS
• GeneAmp ® 5700 SDS
Nonspecific amplification, including primer-dimers, may affect the
quality of amplification data.
Dissociation
curve of
specific product
Dissociation
curve of a
primer dimer
Temperature (°C)
The dissociation curves above show typical primer-dimer formation.
The specific product is shown with a melting temperature (T m ) of
80.5 °C, while the primer-dimer has a characteristically lower T m of
75 °C.
Primer-dimer will be most prevalent in NTC wells and sample wells
containing low concentrations of template.
1-12 Power SYBR
DRAFT
® Green PCR Master Mix Protocol
September 29, 2005 1:41 pm, 1_Chapter.fm