Protocol for flow cytometry-assisted single-nucleus RNA sequencing of human and mouse adipose tissue with sample multiplexing.
STAR Protoc
; 5(1): 102893, 2024 Mar 15.
Article
en En
| MEDLINE
| ID: mdl-38416649
ABSTRACT
Adipocyte size and fragility and commercial kit costs impose significant limitations on single-cell RNA sequencing of adipose tissue. Accordingly, we developed a workflow to isolate and sample-barcode nuclei from individual adipose tissue samples, integrating flow cytometry for quality control, counting, and precise nuclei pooling for direct loading onto the popular 10× Chromium controller. This approach can eliminate batch confounding, and significantly reduces poor-quality nuclei, ambient RNA contamination, and droplet loading-associated reagent waste, resulting in pronounced improvements in information content and cost efficiency.
Palabras clave
Texto completo:
1
Banco de datos:
MEDLINE
Asunto principal:
ARN
/
Núcleo Celular
Límite:
Animals
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Humans
Idioma:
En
Año:
2024
Tipo del documento:
Article