Development of a sequencing system for spatial decoding of DNA barcode molecules at single-molecule resolution.
Commun Biol
; 3(1): 788, 2020 12 18.
Article
em En
| MEDLINE
| ID: mdl-33339962
ABSTRACT
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.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Análise de Sequência de DNA
/
Código de Barras de DNA Taxonômico
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Imagem Individual de Molécula
Limite:
Humans
Idioma:
En
Ano de publicação:
2020
Tipo de documento:
Article