Droplet Digital PCR (ddPCR) Services

At Frontage, we routinely perform accurate, precise, and ultrasensitive quantification ddPCR assays for DNA and RNA to support several applications, including pharmacokinetics (e.g., DNA, mRNA, and oligonucleotide drugs), pharmacodynamics (e.g., gene expression analysis), pharmacogenomics (e.g., mutations and DNA copy number variation), safety (e.g., viral shedding, biodistribution, etc.) and microbial quantification.

What is ddPCR?

ddPCR (droplet digital PCR) is a digital PCR method based on water-oil emulsion droplet technology. A small (~22 μL) reaction mix is fractionated into ~20,000 droplets, such that the vast majority of droplets contain either zero or one copy of the template. PCR amplification occurs in each individual droplet during a standard PCR reaction. After the reaction, each droplet is analyzed to determine whether amplification of the template occurred, and the result is then converted to the template copy number in the original reaction mix using a Poisson Distribution Analysis. ddPCR results thus provide direct quantifications of templates as standard curves are not needed. 

Sample Input Guidelines 

• Extracted DNA or RNA
• Fresh, Frozen or Fixed Cells of Tissues
• Biofluids (e.g., Blood, BM, CSF, Saliva, Tears, Urine, etc)
• Other samples (e.g., Swabs, Fecal Materials, etc)

Advantages of Digital PCR 

• Direct absolute quantification: Standard curves are not needed to convert amplification signals to template copy numbers.  
• Resistant to PCR inhibitors: Compared to qPCR, ddPCR reactions are highly resistant to PCR polymerase inhibitors, thus minimizing matrix effects and providing high-quality results in the presence of other inhibitors. 
• Higher sensitivity: the sensitivity of ddPCR is higher for low copy number templates, providing better lower limit of detection (LLOD) and lower limit of quantification (LLOQ).

Applications of Droplet Digital PCR 

• Pharmacokinetics: quantification of DNA, mRNA, and oligonucleotide pharmaceuticals 
• Pharmacodynamics: gene expression analysis 
• Pharmacogenomics: mutations and DNA copy number variation detection 
• Safety: viral shedding, biodistribution
• Microbial quantification  
• And much more…