Digital PCR technology is a new nucleic acid detection and quantification technology. The Limit of Blank (LOB) is an important parameter for judging the detection capability of digital PCR technology.
LOB (the limit of blank) is a statistical experimental parameter. At a probability of greater than 95% confidence, LOB is the highest possible detection for samples that do not contain analytes (blank samples). In other words, the largest number of false positives.
Why calculate LOB in digital PCR? In digital PCR (dPCR), a variety of factors can cause false positives. Determining false positive cutoffs when quantifying nucleic acids is critical to the stability of dPCR assays. To define this cutoff, the detection threshold is usually set above the LOB value, and a blank LOB value must first be determined for each target during the detection calibration. In addition, the experimental LOB value is necessary to determine whether the reaction is positive or negative and to calculate the LOD value.
The 95% confidence LOB is shown in the following table:
R: number of repetitions, generally ≧30 μ and α: the mean and standard deviation of the number of false positives respectively. μ corr : corrected mean value when μ = 0 (ie false positives never appear), then LOB (95%) = 0
When the LOB is non-zero, the probability distribution weighted "subtract false positive" method can be used to correct the amount of target nucleic acid in the experimental sample.
More LOB values ​​can be found in the Gene-π Digital PCR Academy ( Digital PCR Academy Quick Entry ) statistical tool. You can also choose other application tutorials of interest, including details from experimental design to data analysis. work process. You can also search for specific items directly by searching the navigation tools ("DNA Preparation for dPCR", "Fluorescence Overflow", etc.), or click on our "HOW TO" to select the part you need (design, analysis, report).
LOB (the limit of blank) is a statistical experimental parameter. At a probability of greater than 95% confidence, LOB is the highest possible detection for samples that do not contain analytes (blank samples). In other words, the largest number of false positives.
Why calculate LOB in digital PCR? The 95% confidence LOB is shown in the following table:
R: number of repetitions, generally ≧30 When the LOB is non-zero, the probability distribution weighted "subtract false positive" method can be used to correct the amount of target nucleic acid in the experimental sample.
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