The DOI matches its Crossref record; 30 of the 30 references it cites were found in a registry or an index.
High-Throughput Quantitative Polymerase Chain Reaction in Picoliter Droplets
Open accessAbstract
Limiting dilution PCR has become an increasingly useful technique for the detection and quantification of rare species in a population, but the limit of detection and accuracy of quantification are largely determined by the number of reactions that can be analyzed. Increased throughput may be achieved by reducing the reaction volume and increasing processivity. We have designed a high-throughput microfluidic chip that encapsulates PCR reagents in millions of picoliter droplets in a continuous oil flow. The oil stream conducts the droplets through alternating denaturation and annealing zones, resulting in rapid (55-s cycles) and efficient PCR amplification. Inclusion of fluorescent probes in the PCR reaction mix permits the amplification process to be monitored within individual droplets at specific locations within the microfluidic chip. We show that amplification of a 245-bp adenovirus product can be detected and quantified in 35 min at starting template concentrations as low as 1 template molecule/167 droplets (0.003 pg/microL). The frequencies of positive reactions over a range of template concentrations agree closely with the frequencies predicted by Poisson statistics, demonstrating both the accuracy and sensitivity of this platform for limiting dilution and digital PCR applications.
Overview
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
Integrity signals
Registry
Cites
Statistics
Verification
Claims the paper makes
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
Self-check
Percentages, sums and reported differences re-computed from the paper’s own numbers.
The self-check needs the paper’s full text.
Citations
Cites
Genome sequencing in microfabricated high-density picolitre reactors
Margulies, M., Egholm, M., Altman, W. E., Attiya, S., Bader, J. S., Bemben, L. A., Berka, J., Braverman, M. S., Chen, Y.-J., Chen, Z., et al. (2005) · Nature · doi:10.1038/nature03959 · Cited by 7,693
Self-citationHeid, C. A., Stevens, J., Livak, K. J., & Williams, P. M. (1996) · Genome Research · doi:10.1101/gr.6.10.986 · Cited by 6,174
Rapid Prototyping of Microfluidic Systems in Poly(dimethylsiloxane)
Duffy, D. C., McDonald, J. C., Schueller, O. J. A., & Whitesides, G. M. (1998) · Analytical Chemistry · doi:10.1021/ac980656z · Cited by 5,288
Soft Lithography in Biology and Biochemistry
Whitesides, G. M., Ostuni, E., Takayama, S., Jiang, X., & Ingber, D. E. (2001) · Annual Review of Biomedical Engineering · doi:10.1146/annurev.bioeng.3.1.335 · Cited by 2,607
Formation of droplets and bubbles in a microfluidic T-junction—scaling and mechanism of break-up
Garstecki, P., Fuerstman, M. J., Stone, H. A., & Whitesides, G. M. (2006) · Lab on a Chip · doi:10.1039/b510841a · Cited by 2,339
Formation of dispersions using “flow focusing” in microchannels
Anna, S. L., Bontoux, N., & Stone, H. A. (2003) · Applied Physics Letters · doi:10.1063/1.1537519 · Cited by 2,302
Dynamic Pattern Formation in a Vesicle-Generating Microfluidic Device
Thorsen, T., Roberts, R. W., Arnold, F. H., & Quake, S. R. (2001) · Physical Review Letters · doi:10.1103/physrevlett.86.4163 · Cited by 2,051
Vogelstein, B., & Kinzler, K. W. (1999) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.96.16.9236 · Cited by 1,770
A Microfluidic System for Controlling Reaction Networks in Time
Song, H., Tice, J. D., & Ismagilov, R. F. (2003) · Angewandte Chemie International Edition · doi:10.1002/anie.200390203 · Cited by 1,082
Background and threshold: critical comparison of methods of determination
Reimann, C., Filzmoser, P., & Garrett, R. G. (2005) · Science of The Total Environment · doi:10.1016/j.scitotenv.2004.11.023 · Cited by 886
Cited by
High-throughput droplet digital PCR system for absolute quantitation of DNA copy number
Hindson, B. J., Ness, K. D., Masquelier, D. A., Belgrader, P., Heredia, N. J., Makarewicz, A. J., Bright, I. J., Lucero, M. Y., Hiddessen, A. L., Legler, T. C., et al. (2011) · Analytical Chemistry · doi:10.1021/ac202028g · Cited by 2,792
Self-citationSeemann, R., Brinkmann, M., Pfohl, T., & Herminghaus, S. (2012) · Reports on Progress in Physics · doi:10.1088/0034-4885/75/1/016601 · Cited by 1,049
Quan, P.-L., Sauzade, M., & Brouzes, E. (2018) · Sensors · doi:10.3390/s18041271 · Cited by 664
Surfactants in droplet-based microfluidics
Baret, J.-C. (2012) · Lab Chip · doi:10.1039/c1lc20582j · Cited by 649
Exosome Processing and Characterization Approaches for Research and Technology Development
Lai, J. J., Chau, Z. L., Chen, S., Hill, J. J., Korpany, K. V., Liang, N., Lin, L., Lin, Y., Liu, J. K., Liu, Y., et al. (2022) · Advanced Science · doi:10.1002/advs.202103222 · Cited by 623
Heyries, K. A., Tropini, C., VanInsberghe, M., Doolin, C., Petriv, O. I., Singhal, A., Leung, K., Hughesman, C. B., & Hansen, C. L. (2011) · Nature Methods · doi:10.1038/nmeth.1640 · Cited by 305
Digital Assays Part I: Partitioning Statistics and Digital PCR
Basu, A. S. (2017) · SLAS Technology · doi:10.1177/2472630317705680 · Cited by 300
Mazutis, L., Araghi, A. F., Miller, O. J., Baret, J.-C., Frenz, L., Janoshazi, A., Taly, V., Miller, B. J., Hutchison, J. B., Link, D., et al. (2009) · Analytical Chemistry · doi:10.1021/ac900403z · Cited by 263
Self-citationShen, F., Davydova, E. K., Du, W., Kreutz, J. E., Piepenburg, O., & Ismagilov, R. F. (2011) · Analytical Chemistry · doi:10.1021/ac200247e · Cited by 240
Kreutz, J. E., Munson, T., Huynh, T., Shen, F., Du, W., & Ismagilov, R. F. (2011) · Analytical Chemistry · doi:10.1021/ac201658s · Cited by 148
Benz, C., Retzbach, H., Nagl, S., & Belder, D. (2013) · Lab on a Chip · doi:10.1039/c3lc00057e · Cited by 38
Metadata
- Authors
- Kiss, M. M., Ortoleva-Donnelly, L., Beer, N. R., Warner, J., Bailey, C. G., Colston, B. W., Rothberg, J. M., Link, D. R., & Leamon, J. H.
- Venue
- Analytical Chemistry
- Publisher
- American Chemical Society (ACS)
- Published
- 2008-10-29
- Volume
- 80
- Issue
- 23
- Pages
- 8975-8981
- ISSN
- 0003-2700 (ISSN-L)
- 1520-6882
- OpenAlex
- W2109397680
- Cited by
- 326
- References
- 30
- FWCI
- 17.05
- Type
- journal article
- Language
- en
- Open access
- green
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
Are you an author, or do you see a problem with this page? Request a review
Data sources
Bibliographic data: Semantic Scholar, OpenAlex, Crossref.
Contains information from the Semantic Scholar Academic Graph Datasets, made available under the ODC Attribution License (ODC-BY 1.0).
Semantic Scholar’s data is described in: Kinney, R., et al. (2023). The Semantic Scholar Open Data Platform. arXiv:2301.10140.
