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Digital Droplet PCR (ddPCR) is the 3rd generation of PCR technology, offering ultra-precise, absolute quantification of DNA and RNA. In this video, we break down how ddPCR works, what makes it different from qPCR, and why it's useful in molecular diagnostics, cancer research, and gene expression studies. The polymerase chain reaction (PCR) has undergone modifications through three generations. The first generation of PCR is relies on gel electrophoresis for the analysis of PCR products, yet it faces limitations due to its inadequate detection threshold, labor-intensive procedures, and its singular application scope (qualitative). The subsequent generation of PCR, commonly referred to as real-time quantitative PCR (RT-qPCR), is capable of quantifying products using a standard curve; however, it also exhibits low tolerance to the presence of interfering substances. Digital PCR (dPCR), representing the third generation of PCR, facilitates absolute quantification by partitioning the reaction into discrete compartments. . . . (Information sources: Mao et al., Principles of digital PCR and its applications in current obstetrical and gynecological diseases, Am J Transl Res. 2019; 11(12): 7209–7222. and Thermo fisher Scientific). Quantitation of targets for PCR by use of limiting dilution by P J Sykes 1, S H Neoh, M J Brisco, E Hughes, J Condon, A A Morley https://pubmed.ncbi.nlm.nih.gov/1389177/ . . . . . . . . . This video is about Droplet Digital PCR, ddPCR protocol, Digital PCR technology Quantitative PCR, ddPCR vs. qPCR, DNA quantification methods, Molecular diagnostics Digital PCR applications, High-throughput PCR, Absolute quantification, Rare mutation detection, Copy number variation analysis, Gene expression analysis, Liquid biopsy applications and PCR workflow optimization.