What are the limitations of PCR technology?

Posted June 22, 2020

DNA and RNA Quantitation

Polymerase Chain Reaction (PCR)

Real-Time PCR (qPCR)

Answer

Despite of the widespread application of PCR, this technology has some limitations:

PCR cannot be used to amplify unknown targets. Prior information about the target sequence is necessary to design the primers.
DNA polymerases are prone to error, which potentially causes mutations in PCR products.
PCR is very sensitive to contamination. A small amount of contaminating DNA could result in misleading or ambiguous results.

Additional resources

6-ROX glycine *25 uM fluorescence reference solution for PCR reactions*

Chen, S. C., Halliday, C. L., & Meyer, W. (2002). A review of nucleic acid-based diagnostic tests for systemic mycoses with an emphasis on polymerase chain reaction-based assays. Medical mycology, 40(4), 333-357.

Chuang, L. Y., Cheng, Y. H., & Yang, C. H. (2013). Specific primer design for the polymerase chain reaction. Biotechnology letters, 35(10), 1541-1549.

Related questions

How do hydrolysis probes work in qPCR? probes work?

What are the advantages of Taq polymerase in PCR?

What are the differences between the detection methods based on a fluorescent probe and an intercalating dye in qPCR?

What are the limitations of Taq polymerase?

What are the three stages of a PRC process?