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Benchmarking full-length transcript single cell mRNA sequencing protocols.

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  • Additional Information
    • Source:
      Publisher: BioMed Central Country of Publication: England NLM ID: 100965258 Publication Model: Electronic Cited Medium: Internet ISSN: 1471-2164 (Electronic) Linking ISSN: 14712164 NLM ISO Abbreviation: BMC Genomics Subsets: MEDLINE
    • Publication Information:
      Original Publication: London : BioMed Central, [2000-
    • Subject Terms:
      Benchmarking* ; Transcriptome*; Sequence Analysis, RNA/methods ; High-Throughput Nucleotide Sequencing/methods ; RNA, Messenger/genetics ; Gene Expression Profiling/methods ; Reproducibility of Results ; Single-Cell Analysis
    • Abstract:
      Background: Single cell mRNA sequencing technologies have transformed our understanding of cellular heterogeneity and identity. For sensitive discovery or clinical marker estimation where high transcript capture per cell is needed only plate-based techniques currently offer sufficient resolution.
      Results: Here, we present a performance evaluation of four different plate-based scRNA-seq protocols. Our evaluation is aimed towards applications taxing high gene detection sensitivity, reproducibility between samples, and minimum hands-on time, as is required, for example, in clinical use. We included two commercial kits, NEBNext® Single Cell/ Low Input RNA Library Prep Kit (NEB®), SMART-seq® HT kit (Takara®), and the non-commercial protocols Genome & Transcriptome sequencing (G&T) and SMART-seq3 (SS3). G&T delivered the highest detection of genes per single cell. SS3 presented the highest gene detection per single cell at the lowest price. Takara® kit presented similar high gene detection per single cell, and high reproducibility between samples, but at the absolute highest price. NEB® delivered a lower detection of genes but remains an alternative to more expensive commercial kits.
      Conclusion: For the tested kits we found that ease-of-use came at higher prices. Takara can be selected for its ease-of-use to analyse a few samples, but we recommend the cheaper G&T-seq or SS3 for laboratories where a substantial sample flow can be expected.
      (© 2022. The Author(s).)
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    • Contributed Indexing:
      Keywords: Benchmarking; Full-length RNAseq; G&T sequencing; NEB; SMART-seq3; Single cell; Takara; mRNA sequencing technologies
    • Accession Number:
      0 (RNA, Messenger)
    • Publication Date:
      Date Created: 20221229 Date Completed: 20230102 Latest Revision: 20230103
    • Publication Date:
      20260130
    • Accession Number:
      PMC9801581
    • Accession Number:
      10.1186/s12864-022-09014-5
    • Accession Number:
      36581800