ArticleAnalytical chemistry2014
Digital encoding of cellular mRNAs enabling precise and absolute gene expression measurement by single-molecule counting.
Article in Analytical chemistry, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 14 papers.
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Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
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Who cites it
14 citing papers in PubMed, 30 citations in OpenAlex.
- Transcriptional profiling of circulating tumor cells in multiple myeloma: a new model to understand disease dissemination.Leukemia · 2020Article
- High-Throughput Transcriptome Profiling in Drug and Biomarker Discovery.Frontiers in genetics · 2020Review
- Cell type-dependent differential activation of ERK by oncogenic KRAS in colon cancer and intestinal epithelium.Nature communications · 2019Article
- An integrated enrichment system to facilitate isolation and molecular characterization of single cancer cells from whole blood.Cytometry. Part A : the journal of the International Society for Analytical Cytology · 2018Article
- High-throughput determination of the antigen specificities of T cell receptors in single cells.Nature biotechnology · 2018Article
- Elimination of PCR duplicates in RNA-seq and small RNA-seq using unique molecular identifiers.BMC genomics · 2018Article
- High-throughput sequencing of murine immunoglobulin heavy chain repertoires using single side unique molecular identifiers on an Ion Torrent PGM.Oncotarget · 2018Article
- Cholesterol signaling in single cells: lessons from STAR and sm-FISH.Journal of molecular endocrinology · 2018Review
- An integrated flow cytometry-based platform for isolation and molecular characterization of circulating tumor single cells and clusters.Scientific reports · 2018Article
- Immune Repertoire Sequencing Using Molecular Identifiers Enables Accurate Clonality Discovery and Clone Size Quantification.Frontiers in immunology · 2018Article
- RNA-Seq methods for transcriptome analysis.Wiley interdisciplinary reviews. RNA · 2017Review
- Single-cell analysis tools for drug discovery and development.Nature reviews. Drug discovery · 2016Review
- Accurate and predictive antibody repertoire profiling by molecular amplification fingerprinting.Science advances · 2016Article
- Single-cell transcriptome sequencing: recent advances and remaining challenges.F1000Research · 2016Review
Corrections and comments
PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.
Authors and funding
5 authors at 1 institution in 1 country.
Funding
Abstract
We present a new approach for the sensitive detection and accurate quantitation of messenger ribonucleic acid (mRNA) gene transcripts in single cells. First, the entire population of mRNAs is encoded with molecular barcodes during reverse transcription. After amplification of the gene targets of interest, molecular barcodes are counted by sequencing or scored on a simple hybridization detector to reveal the number of molecules in the starting sample. Since absolute quantities are measured, calibration to standards is unnecessary, and many of the relative quantitation challenges such as polymerase chain reaction (PCR) bias are avoided. We apply the method to gene expression analysis of minute sample quantities and demonstrate precise measurements with sensitivity down to sub single-cell levels. The method is an easy, single-tube, end point assay utilizing standard thermal cyclers and PCR reagents. Accurate and precise measurements are obtained without any need for cycle-to-cycle intensity-based real-time monitoring or physical partitioning into multiple reactions (e.g., digital PCR). Further, since all mRNA molecules are encoded with molecular barcodes, amplification can be used to generate more material for multiple measurements and technical replicates can be carried out on limited samples. The method is particularly useful for small sample quantities, such as single-cell experiments. Digital encoding of cellular content preserves true abundance levels and overcomes distortions introduced by amplification.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.