Seven Digital PCR Features You're Probably Ignoring
Seven Digital PCR Features You're Probably Ignoring Pixabay

Nucleic acid amplification has been the choice technique for detecting genetic markers for viruses and bacteria for years. Its ability to pick up RNA and DNA instead of antigens has made it one of the most sensitive technologies in the industry, but digital polymerase chain reaction (digital PCR) is taking it to new heights. Polymerase chain reactions can make millions of DNA copies, so scientists need only a tiny sample to identify infectious agents. Today, genetic treasure hunts are more powerful than ever before, so researchers can detect even the faintest signals. As long as you can find one positive in hundreds of thousands of negatives, you can find that one all-important marker that changes the game. Most PCR techniques use thermal cycling to achieve nucleic acid denaturation. Once DNA stands have been separated, temperatures can be reduced and primers can be bonded. These are used as templates that ultimately generate DNA themselves, creating a chain reaction. Digital PCR can be used in cloning, mutagenesis, and genetic analysis, so it's made PCR infinitely easier to achieve. The technique might be powerful, but it is only as effective as the scientist that wields it. Here are a few dPCR truths you're probably ignoring.

1) It Relies on Data Sharing

Digital PCR must be quantitative despite its qualitative elements, so it charts the former as it performs the latter. The process uses up an incredible amount of genetic data, so if you don't have an adequate data sharing tool, you're unlikely to enjoy much success. You need excellent workflow and data analysis tools to benefit.

2) It's Only as Powerful as Its Partitioning Method

Digital PCR and partitioning statistics must work hand in hand. dPCR copes with dynamic range and big sample volumes by comparing binary outputs. Partitioning can improve the purification of one of two replicates, which in turn makes amplification more efficient. The more partitions you have, the more precise you will be.

3) It Can Achieve More Precision than Real-Time PCR

Quantitative PCR has been the diagnostics niche's favourite workhorse for years, but digital PCR has performed better in recent trials. It also delivers more reproducible results without losing any sensitivity, so its role in diagnosing HIV is secured. That's one of many reasons that Stilla Technologies favours it for HIV provirus filing. Not only does it achieve more accuracy, but also improved detection of gene expression.

4) It Can Reduce Operating Costs

With the right system workflow, dPCR can be as economical as it is accurate and quick. The naica® systemworkflow, for example, can handle up to 20, 000 droplets per sample and 16 samples per Opal chip. Once that's done, PCR amplification can follow , and all that's required is a single pipetting operation.

5) The Right Master Mix PCR Protocol Can Improve Results

Your PCR reaction is only as efficient as the quality of your PCR mixture components. It should be optimised to inactivate enzyme activity until heat is applied and the PCR cycle is triggered. This can improve both sensitivity and specificity. Add PCR-grade water, a good primer, and a solid master mix PCR protocol to achieve the best results.

6) Digital PCR Assays Are Easier to Achieve than You Might Think

When digital PCR was first developed, the process had to be handled manually. Today, PCR assays can be achieved with a protocol that's even more efficient than 2003's BEAM. Hundreds of thousands of wells can be separated in huge quantities. With the right platform, next generation libraries can perform quality control prior to sequencing. New protocols can cope with loci of more than 30, and parallel sequencing can work with hundreds of millions of template molecules.

7) Digital PCR Has Important Cancer-Related Benefits

dPCR's sheer breadth has made it a core technology for several fields, from farming and fetal gender determination to genotyping. It's become an important way to evaluate archival tumor tissues, so oncology has become less invasive. It's also opened up new diagnostic and predictive tools to spot malignancies.

Automation and cost-efficiency tend to coexist for obvious reasons. Computers don't need salaries, so the more multiplexing you can build into the dPCR process, the cheaper it becomes. Today, two four-plex assays can be developed for every eight DNA targets. As computing evolves, the process will become even more powerful. Digital PCR has a massive breadth of relevancies, from everyday cancer diagnoses to complex GMO crop events. With so many PCR replicates on hand, precision never needs to be sacrificed.