What are the differences between real time PCR and nested PCR?

Question

Accepted Answer

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

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