Introduction
The amplification of DNA samples before their sequencing is a crucial step that has become routine. The isothermal amplification of nucleic acids represents a simplified process that allows for the quick and efficient accumulation of nucleic acid sequences in an environment of constant temperatures.
Discussion
Since the beginning of the 1990s, different approaches to isothermal amplification have emerged as alternative methods to PCR (Zanoli & Spoto, 2012). This technique has been implemented for biosensing targets which include DNA, RNA, various cells, as well as proteins, ions, and small molecules. The core difference between PCR and isothermal amplification lies in the temperature changes. When it comes to PCR, a thermal cycler facilitates a shift in reaction temperatures repeatedly to affect the actions of temperature-reliant reagents. However, isothermal amplification takes place in an environment of a single temperature. Such a method provides the ability to amplify DNA at a static temperature and use strand-displacing polymerases of DNA for “unzipping” strands as they go through double-stranded DNA instead of using heat denatures as in PCR; this allows for primers’ access to templates.
Conclusion
Overall, choosing either PCR or isothermal amplification varies depending on the individual application and the equipment that is available. Without a thermal cycler, PCR cannot be implemented, which could be a disadvantage for diagnostic applications and point-of-care settings. Isothermal amplification is perfect for situations in which a thermal cycler is unavailable, such as clinics or home testing kits. This method can be quicker, less complicated, and more cost-effective. To conclude, there could be various reasons for using either method for the amplification of DNA samples, which is why it is important to note their differences in usage.
References
Zanoli, L. M., & Spoto, G. (2012). Isothermal amplification methods for the detection of nucleic acids in microfluidic devices. Biosensors, 3(1), 18–43. Web.