Digital droplet PCR (ddPCR) is a PCR-based technology that employs water-oil immersion droplet technology to partition study samples containing template DNA molecules. Each droplet is essentially an individual well in a plate or test tube in which a routine PCR reaction takes place. This sample partitioning is a critical element of ddPCR assays.
ddPCR often partitions nucleic acid study samples into thousands of droplets, where PCR amplification takes place within each nanometer-sized droplet. This method requires smaller sample volumes and reagents than other PCR systems, preserving precious biological samples and reducing associated assay costs. The current article dives deep into ddPCR gene expression analysis and provides an understanding of how ddPCR assays help unravel the genetic activity and provide deep insights.
Understanding genetic activity and gene expression analysis with ddPCR services
Partitioning study samples helps researchers detect single DNA templates with precision, sensitivity, and specificity. It also reduces target competition effects, making product amplification robust against inhibition while improving the discriminatory ability of ddPCR assays to detect single nucleotides. ddPCR gene expression analysis has several benefits and is preferred in multiple therapeutic areas.
ddPCR assays can quantify target DNA in the samples without requiring standard curves. Hence, this technique is ideal for viral load analysis, microbial quantification, and target DNA measurement.
ddPCR is an ideal copy number variation assay. Traditionally, qPCR copy number assays were widely employed by researchers and scientists. Copy number variations cause too many or too few dosage-dependent changes influencing complex behavioral traits, phenotypic variability, and diseases. ddPCR assays offer measurements with 1.2X differences in copy numbers.
To better understand disease biology, researchers amplify single genes such as T-cells present in wild-type backgrounds. ddPCR assays are sensitive and robust to detect rare sequences or mutations in these complex samples.
ddPCR is ideal for microRNA and gene expression analysis. They offer stand-alone quantification results of expression levels, especially microRNAs present in low abundance with precision and sensitivity.
ddPCR assays are an ideal partner for next-generation sequencing. They can quantify next-generation sequencing sample library preparations to reduce run repeats and increase sequencing accuracy. Besides, ddPCR assays can validate results such as copy number variations and single nucleotide polymorphism with absolute quantification.
Single-cell analysis is critical for gene expression studies. The primary reason for this need is the high degree of cell-cell variation observed in genomic and gene expression content among progenitor homogenous post-mitotic and stem cell populations. ddPCR assays can effectively quantify low copy numbers present in complex study material.
Importantly, advanced genome editing tools require robust and accurate assessment and quantification of edited products. ddPCR can precisely assess homologous directed repair and nonhomologous end products generated by genome editing tools such as Crispr-cas9.
In Conclusion
ddPCR assays can provide deep insights into genetic activity through gene expression analysis. However, similar to bioanalytical assessments, such as (PK) pharmacokinetics studies and pharmacodynamic (PD) assays, ddPCR gene expression analysis will require a collaborative effort to expand its applications in different biomedical and clinical research domains.
