Development of a sequencing system for spatial decoding of DNA barcode molecules at single-molecule resolution.
Journal
Communications biology
ISSN: 2399-3642
Titre abrégé: Commun Biol
Pays: England
ID NLM: 101719179
Informations de publication
Date de publication:
18 12 2020
18 12 2020
Historique:
received:
11
05
2020
accepted:
17
11
2020
entrez:
19
12
2020
pubmed:
20
12
2020
medline:
22
6
2021
Statut:
epublish
Résumé
Single-cell transcriptome analysis has been revolutionized by DNA barcodes that index cDNA libraries, allowing highly multiplexed analyses to be performed. Furthermore, DNA barcodes are being leveraged for spatial transcriptomes. Although spatial resolution relies on methods used to decode DNA barcodes, achieving single-molecule decoding remains a challenge. Here, we developed an in-house sequencing system inspired by a single-molecule sequencing system, HeliScope, to spatially decode DNA barcode molecules at single-molecule resolution. We benchmarked our system with 30 types of DNA barcode molecules and obtained an average read length of ~20 nt with an error rate of less than 5% per nucleotide, which was sufficient to spatially identify them. Additionally, we spatially identified DNA barcode molecules bound to antibodies at single-molecule resolution. Leveraging this, we devised a method, termed "molecular foot printing", showing potential for applying our system not only to spatial transcriptomics, but also to spatial proteomics.
Identifiants
pubmed: 33339962
doi: 10.1038/s42003-020-01499-8
pii: 10.1038/s42003-020-01499-8
pmc: PMC7749132
doi:
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
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