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