Exact(6)
In [ 38], the authors have developed AgentCell, an ABM based digital assay for the study of bacterial chemotaxis.
In the first approach, we demonstrate that direct quantification using the digital assay compares well to a qPCR standard curve, validating the general performance of the assay.
We detected no digital PCR counts in a mock (no sample) 454 library preparation [see Additional file 1], indicating that the digital assay is a background-free measurement.
Because this digital assay does not rely on internal or external standards, the RSD of approximately 3.0% demonstrates the true accuracy of the assay.
Because the digital assay relies on neither internal nor external standards, the CV figure of 11.8% represented in Figure 3B closely approximates the real-world accuracy of the digital assay, which is sufficient to prepare bulk emulsion PCR or bridge PCR reactions without prior titration.
In the second approach, we compare the direct quantification of single-cell mRNA from the digital assay to another direct RNA counting method, single-molecule FISH (fluorescence in situ hybridization), indicating the accuracy of postdigitization RT.
Similar(54)
We explain how to design classic digital assays, comprising identical partitions, in order to obtain the required precision of the estimate within a defined range of concentrations.
The UT-digital assay is performed on a commercial microfluidic digital PCR chip from Fluidigm.
In this study, we performed the EGG analysis only in pregnancies involving male fetuses because a precise digital PCR assay targeting the Y-chromosome, namely the ZFY DNA assay, is well established in our laboratory [5].
Using a novel droplet digital PCR assay and high-throughput sequencing, we characterized the frequency of rearrangements between the β- and δ-globin paralogs when delivering these nucleases.
β-actin DNA was essentially undetectable by the digital PCR assay in all of the 36 digested maternal plasma DNA samples, implying that complete digestion had occurred.
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