Basis of differentiation | Real time PCR | Nested PCR |
Definition | In real-time PCR, the amplification product is measured and quantified as the reaction progresses, providing continuous monitoring of the process | Nested PCR is a method used to minimize nonspecific amplification of DNA templates and involves two successive amplification reactions |
Two successive amplification reactions | Does not involve two successive amplification reactions | Involves two successive amplification reactions |
Quantification of the product | Yes | No since it is typically used for qualitative analysis |
Gel-electrophoresis requirement | No since it monitors fluorescence to observe amplification | Yes |
Use of two sets of primers | No | Yes |
Reducing non-specific amplification | No | Yes |
Time required | Requires less time since | Requires more time due to multiple PCR cycles |
Risk of contamination | Higher risk due to frequent opening of tubes for sampling | Lower risk as nested amplification occurs in a closed system |