Theoretical evaluation of the probabilistic characteristics of a quantitative analysis based on a combination of droplet microfluidic and polymerase chain reaction for extremely small amounts of genetic material

The classical method of real-time polymerase chain reaction (PCR) has a large error in determining ultra-small amounts of genetic material (hundreds copies or less). In these conditions, the methods of digital PCR are used. At the same time, the smallest error in quantitative analysis is achieved under conditions of comparability of the number of copies and the number of analytical cells in which the presence of at least one copy of the analyzed material is recorded. The paper presents a theoretical probabilistic calculation of a scheme of a principal new combination of PCR and droplet microfluidic, which will provide acceptable characteristics for the analysis of extremely small amounts of the sample. In this case, PCR is used exclusively for the copying of genetic material, and quantitative analysis is performed with the help of droplet microfluidic. An example of estimating the error in counting the number of copies is given, taking into account two different independent random factors, as that selection of 10% of the sample and 1% error of drops’ with positive results of reaction estimation.