ArticlePloS one2011
Decoding a substantial set of samples in parallel by massive sequencing.
Article in PloS one, 2011. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 10 papers.
What it found
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The trial behind it
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
10 citing papers in PubMed, 25 citations in OpenAlex.
- An integrative protocol for one-step PCR amplicon library construction and accurate demultiplexing of pooled sequencing data.Marine life science & technology · 2023Article
- Statistical assessment of DNA extraction methodology for culture-independent analysis of microbial community associated with diverse environmental samples.Molecular biology reports · 2018Article
- An automated approach to prepare tissue-derived spatially barcoded RNA-sequencing libraries.Scientific reports · 2016Article
- Next-generation genotyping of hypervariable loci in many individuals of a non-model species: technical and theoretical implications.BMC genomics · 2016Article
- Phasing of single DNA molecules by massively parallel barcoding.Nature communications · 2015Article
- Hierarchical molecular tagging to resolve long continuous sequences by massively parallel sequencing.Scientific reports · 2013Article
- Targeted transcript profiling by sequencing.Scientific reports · 2012Article
- Library preparation and multiplex capture for massive parallel sequencing applications made efficient and easy.PloS one · 2012Article
- High-throughput multilocus sequence typing: bringing molecular typing to the next level.PloS one · 2012Article
- Rapid flow-sorting to simultaneously resolve multiplex massively parallel sequencing products.Scientific reports · 2011Article
Corrections and comments
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Authors and funding
4 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
There has been a dramatic increase of throughput of sequenced bases in the last years but sequencing a multitude of samples in parallel has not yet developed equally. Here we present a novel strategy where the combination of two tags is used to link sequencing reads back to their origins from a pool of samples. By incorporating the tags in two steps sample-handling complexity is lowered by nearly 100 times compared to conventional indexing protocols. In addition, the method described here enables accurate identification and typing of thousands of samples in parallel. In this study the system was designed to test 4992 samples using only 122 tags. To prove the concept of the two-tagging method, the highly polymorphic 2(nd) exon of DLA-DRB1 in dogs and wolves was sequenced using the 454 GS FLX Titanium Chemistry. By requiring a minimum sequence depth of 20 reads per sample, 94% of the successfully amplified samples were genotyped. In addition, the method allowed digital detection of chimeric fragments. These results demonstrate that it is possible to sequence thousands of samples in parallel without complex pooling patterns or primer combinations. Furthermore, the method is highly scalable as only a limited number of additional tags leads to substantial increase of the sample size.
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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.