The Power of Partitioning: The World of Droplet Digital™ PCR Applications

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Since its introduction two years ago, Droplet Digital PCR has transformed multiple fields of research, including infectious disease, cancer biomarker analysis, and genomic variation analysis. This webinar introduces the concept, workflow, and performance of Droplet Digital PCR and highlights the diverse applications in showcased in recent journal publications.
The QX200™ Droplet Digital PCR system, Bio-Rad's second-generation digital PCR system, provides absolute quantification of target DNA or RNA molecules using EvaGreen or TaqMan hydrolysis probes, yielding unmatched sensitivity and precision for a wide variety of applications.
Presenter: Dr. George Karlin-Neumann, Director of Scientific Affairs, Bio-Rad Digital Biology Center
Dr. Karlin-Neumann received his PhD in molecular genetics in 1990 from UCLA. After postdoctoral studies with Dr. Ronald Davis in Stanford's Biochemistry Department, he left in 2001 to cofound ParAllele BioScience, which was acquired by Affymetrix in 2005. Dr. Karlin-Neumann joined QuantaLife in 2010. He has since transitioned to his current role as Director of Scientific Affairs at the Digital Biology Center at Bio-Rad Laboratories, where he is responsible for establishing and over-seeing research collaborations exploring new directions for digital PCR technology.
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[@ShakespeareCafe]

Digital PCR: Methods and Protocols Ed. George Karlin-Neumann and Francisco Bizouarn. Springer Press

[@purushothaman6241]

Very nice

[@haseebzubair3681]

At 17:43 the speaker talks about 20,000 droplets but if I am correct this image shows only 15,000. My question then is, what is the acceptable number of droplets read for a confident result interpretation?

[@IOffspringI]

The more droplets you have, the smaller the confidence interval. There is a formula for the size of the confidence interval as a function of the fraciton of positives and the number of partitions (droplets) given in
Michael Rabe, Dorinel Verdes, and Stefan Seeger. Understanding protein adsorptionphenomena at solid surfaces.Advances in Colloid and Interface Science, 162(1-2):87-106, 2011
Personally I would say that everything over 10 000 partitions has sufficiently good results. Hope that helps you.

[@thisismattness3215]

Dear Mr Zubair,
I'm a technician in Germany and i've been working constantly with the QX200 ddPCR system for over a year now.
The 15,000 droplets shown in the graph were probably due to the loss of droplets while pipetting when you use the manual droplet generator.
In my lab we usually aim for an accepted droplet count of 16,000-18,000 and train people as such. We do not evaluate samples below 14,000 accepted droplets and repeat the ddPCR for those sampples. If you want more constant droplet counts, i would advise you to invest in the automated droplet generator. Hope this helped,
Best.

[@sayatamvan5580]

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