Uzbas, F. ; Opperer, F. ; Sönmezer, C. ; Shaposhnikov, D. ; Sass, S. ; Krendl, C. ; Angerer, P. ; Theis, F.J. ; Müller, N.S. ; Drukker, M.
BART-Seq: cost-effective massively parallelized targeted sequencing for genomics, transcriptomics, and single-cell analysis.
Genome Biol. 20:155 (2019)
We describe a highly sensitive, quantitative, and inexpensive technique for targeted sequencing of transcript cohorts or genomic regions from thousands of bulk samples or single cells in parallel. Multiplexing is based on a simple method that produces extensive matrices of diverse DNA barcodes attached to invariant primer sets, which are all pre-selected and optimized in silico. By applying the matrices in a novel workflow named Barcode Assembly foR Targeted Sequencing (BART-Seq), we analyze developmental states of thousands of single human pluripotent stem cells, either in different maintenance media or upon Wnt/beta-catenin pathway activation, which identifies the mechanisms of differentiation induction. Moreover, we apply BART-Seq to the genetic screening of breast cancer patients and identify BRCA mutations with very high precision. The processing of thousands of samples and dynamic range measurements that outperform global transcriptomics techniques makes BART-Seq first targeted sequencing technique suitable for numerous research applications.
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Publication type
Article: Journal article
Document type
Scientific Article
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Keywords
Barcoding ; Single-cell Rna Sequencing ; Targeted Transcriptomics ; High-throughput Screening ; Human Pluripotent Stem Cells ; Multiplex Pcr; Stem-cell; Gene-expression; Differentiation; Brca2; State; Rna
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Language
english
Publication Year
2019
Prepublished in Year
HGF-reported in Year
2019
ISSN (print) / ISBN
1474-760X
e-ISSN
1465-6906
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Article Number: 155
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BioMed Central
Publishing Place
Campus, 4 Crinan St, London N1 9xw, England
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Reviewing status
Peer reviewed
POF-Topic(s)
30204 - Cell Programming and Repair
30205 - Bioengineering and Digital Health
30504 - Mechanisms of Genetic and Environmental Influences on Health and Disease
Research field(s)
Stem Cell and Neuroscience
Enabling and Novel Technologies
PSP Element(s)
G-500800-001
G-503800-001
G-552400-001
G-508200-014
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Erfassungsdatum
2019-08-13