Polymerase chain reaction, application of PCR in clinical diagnostics, RT-PCR and use of this technique.

Polymerase Chain reaction is method of amplifying specific sequences of DNA. It is a method that popular used in the field of medicine due to sensitivity, specificity and how quickly it yields result.

Mechanisms of PCR

Three main steps of PCR:

• Denaturation : DNA is denatured at a temperature of 90 degrees celsius. The double stranded helical DNA is separated into two separated strands
  • This is performed at the temperature of 90 degree celsius
  • The separated stands of DNA serve as template for amplification.
• Annealing : Primers and deoxynucleoside triphosphate are added in excess to the sample of DNA, where in they anneal to the complementary sequences on the strands DNA
  • Primer : a short oligonucleomers that is complementary to the target sequence that needs to the amplified
  • To prevent the quantity of primer and deoxynucleoside limiting the process of PCR, they are added in excess.

References

Rodwell, V.W. et al. (2018) Harper’s Illustrated Biochemistry 31. 31st edn. New York: McGraw-Hill Medical. Pg 1071 - 1074

Polymerase chain reaction (PCR) (n.d) Khan Academy. Available at: https://www.khanacademy.org/science/ap-biology/gene-expression-and-regulation/biotechnology/a/polymerase-chain-reaction-pcr#:~:text=PCR%20is%20used%20in%20many%20research%20labs%2C%20and%20it%20also,the%20case%20of%20prenatal%20testing). (Accessed: 02 April 2025).

Khehra, N., Padda, I.S. and Swift., C.J. (2023) Polymerase chain reaction (PCR), StatPearls [Internet]. Available at: https://www.ncbi.nlm.nih.gov/books/NBK589663/ (Accessed: 02 April 2025).

PCR technique and its applications in biology and medicine (2023) MyBioSource Learning Center. Available at: https://www.mybiosource.com/learn/pcr-technique-and-its-applications-in-biology-and-medicine/ (Accessed: 02 April 2025).

Murray, P.R., Rosenthal, K.S. and Pfaller, M.A. (2016) Medical microbiology. 8th edition. Philadelphia, PA: Elsevier. Page 23 - 25