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What are the limitations of PCR technology?
Posted June 22, 2020
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.