2. Contents
Introduction
Mechanism of Real-time PCR
Types of Real-time PCR probes detection
Advantages & disadvantages of SYBR Green
Advantages & disadvantages of Taqman probe
Advantages of Real-time PCR
Disadvantages of Real time PCR
Applications
3. Introduction
A real-time polymerase chain reaction (real-time PCR) is
a laboratory technique of molecular biology based on
the polymerase chain reaction (PCR).
It monitors the amplification of a targeted DNA molecule
during the PCR (i.e., in real time), not at its end, as in
conventional PCR.
• It is also known as a quantitative polymerase chain reaction
(qPCR)
13. Advantages of rt- PCR:
• High technical sensitivity (<5copies)
• High precision
• No post-PCR steps like running of gel or sequencing etc.
• Non – specific amplification can be detected .
• Minimized risk of cross contamination.
• Saves lot of time.
14. Disadvantages of rt- PCR
• PCR product increases exponentially cannot monitor amplicon
size.
• Can give incorrect quantity if DNA sample is degraded.
• Maximum of four simultaneous reactions.
• Non- specific binding (SYBR Green analysis).
15. Applications:
• Diagnostic uses
• Microbiological uses
• Detection of phytopathogens
• Detection of genetically modified organisms
• Clinical quantification and genotyping