Parnad Basu, Amity University Kolkata
PCR (Polymerase Chain Reaction) is a technique used to amplify small segments of any DNA. In conventional PCR, DNA libraries suffer from template mispairing. This ultimately leads to the obvious loss of unique sequences. To be more efficient, ePCR (emulsion Polymerase Chain Reaction) can be used as it has a fundamental advantage over conventional PCR by deep sequencing and mathematical modelling.
In a recent study, the advantage of using water-in-oil emulsions, PCR opened a new door in terms of DNA amplification. Different types of DNA libraries (random DNA libraries, antibody libraries, etc.) have benefited from ePCR based approaches. In the case of estimation of the NGS (next-generation sequencing) the improvement due to the use of ePCR was very significant. The ePCR resulted in a more uniform distribution of amplicons while improving the yield of amplified DNA libraries. It also reduced the error count during amplification.
The ePCR protocol can be done by either vortexing or magnetic stirring. Between these two, vortexing showed a more uniform distribution of droplet sizes. In the case of magnetic stirring, it shows a decrease in the mean diameter of the droplets. These techniques make ePCR much more approachable, cost-effective, and user friendly concerning the conventional PCR. In a side by side comparison, ePCR overtook conventional PCR in less frequent deletion and gross errors. Not only that, but the mathematical model of an ePCR also helps it to work efficiently. Also, total genetic information can be revealed using quasi single-molecule ePCR which provides the way to preserve the quality of DNA libraries during amplification.
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Reference:- https://doi.org/10.1073/pnas.2017138117
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