Accurate bulk quantitation of droplet digital PCR
Sun, C.; Liu, L.; Vasudevan, H.; Chang, K.-C.; Abate, A.
Show abstract
Droplet digital PCR provides superior accuracy in nucleic acid quantitation. The requirement of microfluidics to generate and analyze the emulsions, however, is a barrier to its adoption, particularly in low resource or clinical settings. Here, we report a novel method to prepare ddPCR droplets by vortexing and readout the results by bulk analysis of recovered amplicons. We demonstrate the approach by accurately quantitating SARS-CoV-2 sequences using entirely bulk processing and no microfluidics. Our approach for quantitating reactions should extend to all digital assays that generate amplicons, including digital PCR and LAMP conducted in droplets, microchambers, or nanoliter wells. More broadly, our approach combines important attributes of ddPCR, including enhanced accuracy and robustness to inhibition, with the high-volume sample processing ability of quantitative PCR.
Matching journals
The top 8 journals account for 50% of the predicted probability mass.
Similar papers in this journal
- Portable real-time colorimetric LAMP-device for rapidquantitative detection of nucleic acids in crude samples 95%
- Digital droplet PCR accurately quantifies SARS-CoV-2 viral load from crude lysate without nucleic acid purification 94%
- Absolute quantification of rare gene targets in limited samples using crude lysate and ddPCR 94%
Similar papers in this journal
Similar papers in this journal
- Keeping it clean: the cell culture quality control experience at the National Center for Advancing Translational Sciences 90%
- waveRAPID(R) - a robust assay for high-throughput kinetic screens with the Creoptix(R) WAVEsystem 89%
- Quantitative target engagement of RIPK1 in human whole blood via the cellular thermal shift assay for potential pre-clinical and clinical applications 88%
Similar papers in this journal
- Development and Implementation of a scalable and versatile test for COVID-19 diagnostics in rural communities 95%
- Laboratory validation of a clinical metagenomic next-generation sequencing assay for respiratory virus detection and discovery 93%
- All-in-One Dual CRISPR-Cas12a (AIOD-CRISPR) Assay: A Case for Rapid, Ultrasensitive and Visual Detection of Novel Coronavirus SARS-CoV-2 and HIV virus 93%
Similar papers in this journal
- Quantitative Isothermal Amplification on Paper Membranes using Amplification Nucleation Site Analysis 96%
- Point-of-care detection of SARS-CoV-2 in nasopharyngeal swab samples using an integrated smartphone-based centrifugal microfluidic platform 95%
- SNAPflex: a paper-and-plastic device for instrument-free RNA and DNA extraction from whole blood 94%
"Similar papers" are the closest papers from that journal in the model's embedding space. They show what the match is built on, but the ranking comes mostly from a classifier over the whole training set, not from these examples alone.