Polymerase Chain Reaction Essay Example
The process of Polymerase Chain Reaction is relatively straightforward. The strand of DNA is first heated up to between 93℃ and 96℃. The heat denatures the DNA sample, essentially splitting the helix into two separate strands (Kubista et al. 2006). These two strands act as templates for the polymerase enzyme to bind the new primers to. In order for the enzyme to work, the temperature must be lowered. While the Taq Polymerase can survive the high temperatures required to denature the DNA it is best utilized at a lower temperature. The perfect temperature for this Taq Polymerase is about 72°C (Kubista et al. 2006). These primers mirror the original strands to create another copy of the original strand. This process of denaturing,priming, and transcribing is then repeated millions of times until the appropriate amount of DNA sample is synthesized.
The DNA Polymerase Reaction began in 1983 with Kary Mullis, a biochemist who was trying to figure out a way to replicate DNA in a repeatable, efficient fashion . At this point, the DNA replications were lackluster at best. The E. Coli Polymerase would become inactive during the denaturing of the DNA, leading to chemists having to add more Polymerase or having very limited products (Bartlett & Stirling 2003). Mullis tackled this problem by utilizing the enzyme called “Taq Polymerase” which could survive the high temperatures utilized when denaturing the DNA samples (Kubista et al. 2006). By utilizing this enzyme, it creates a replicable process that is efficient in its usage. This groundbreaking efficiency gives biochemists millions of copies of the DNA strands to experiment on and work with. This huge change in how DNA replication was done led to Kary Mullis receiving the Nobel Prize for Chemistry in 1993 (Bartlett & Stirling 2003). Without this achievement, we would not be able to replicate DNA at such a rate, slowing down future development in genetic research and understanding.
According to the essay “The Future of Digital Polymerase Chain Reaction in Virology”, digital polymerase chain reaction, or dPCR, has a lot of potential in the future of biology and more specifically virology. Currently dPCR is used to minimize pre-processing of the sequences as well as minimize loss of sequences. It also has been used to detect low levels of contamination that other methods cannot detect. dPCR is also more accurate than other PCR methods. This allows for virologists to have a pretty good idea of what to expect when dealing with strands that could be potentially useful, without having to question whether the results are correct or not.
Less possible variables means faster processing and understanding of the sequences. This efficiency is obviously very useful and can breed great results. Another benefit of using dPCR is its precision and power in use. It is, reportedly, more precise than other PCR methods and does a better job of getting consistent results. This precision along with its accuracy leads to a very reliable source of information that can be utilized much more effectively than it is now.
However, this does not mean that this method is perfect, the standard dPCR equation used to calculate a concentration uses volume, positive partitions, and total partitions. These variables are potentially biased and should be studied further, according to Dr. Vynck. Furthermore, while dPCR is certainly not perfect, with more attention and improvement it can lead to a great many benefits for virologists and biologists everywhere(Vynck, Trypsteen, Thas, Vandekerckhove, & De Spiegelaere, 2016).