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Protocol for flow cytometry-assisted single-nucleus RNA sequencing of human and mouse adipose tissue with sample multiplexing.
Gulko, Anton; Essene, Adam; Belmont-Rausch, Dylan Matthew; Veregge, Molly; Pant, Deepti; Tenen, Danielle; Kapel, Benedicte Schultz; Emont, Margo P; Pers, Tune H; Rosen, Evan D; Tsai, Linus T.
Afiliação
  • Gulko A; Division of Endocrinology, Diabetes, and Metabolism, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA 02115, USA; Boston Nutrition and Obesity Research Center/Boston Area Diabetes and Endocrinology Research Center Functional Genomics and Bioinformatics Core, Boston, MA, USA.
  • Essene A; Division of Endocrinology, Diabetes, and Metabolism, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA 02115, USA; Boston Nutrition and Obesity Research Center/Boston Area Diabetes and Endocrinology Research Center Functional Genomics and Bioinformatics Core, Boston, MA, USA.
  • Belmont-Rausch DM; Novo Nordisk Foundation Center for Basic Metabolic Research, University of Copenhagen, 2200 Copenhagen, Denmark.
  • Veregge M; Division of Endocrinology, Diabetes, and Metabolism, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA 02115, USA; Boston Nutrition and Obesity Research Center/Boston Area Diabetes and Endocrinology Research Center Functional Genomics and Bioinformatics Core, Boston, MA, USA.
  • Pant D; Division of Endocrinology, Diabetes, and Metabolism, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA 02115, USA; Boston Nutrition and Obesity Research Center/Boston Area Diabetes and Endocrinology Research Center Functional Genomics and Bioinformatics Core, Boston, MA, USA.
  • Tenen D; Division of Endocrinology, Diabetes, and Metabolism, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA 02115, USA; Boston Nutrition and Obesity Research Center/Boston Area Diabetes and Endocrinology Research Center Functional Genomics and Bioinformatics Core, Boston, MA, USA.
  • Kapel BS; Novo Nordisk Foundation Center for Basic Metabolic Research, University of Copenhagen, 2200 Copenhagen, Denmark.
  • Emont MP; Division of Endocrinology, Diabetes, and Metabolism, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA 02115, USA.
  • Pers TH; Novo Nordisk Foundation Center for Basic Metabolic Research, University of Copenhagen, 2200 Copenhagen, Denmark.
  • Rosen ED; Division of Endocrinology, Diabetes, and Metabolism, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA 02115, USA; Boston Nutrition and Obesity Research Center/Boston Area Diabetes and Endocrinology Research Center Functional Genomics and Bioinformatics Core, Boston, MA, USA.
  • Tsai LT; Division of Endocrinology, Diabetes, and Metabolism, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA 02115, USA; Boston Nutrition and Obesity Research Center/Boston Area Diabetes and Endocrinology Research Center Functional Genomics and Bioinformatics Core, Boston, MA, USA.
STAR Protoc ; 5(1): 102893, 2024 Mar 15.
Article em 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.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: RNA / Núcleo Celular Limite: Animals / Humans Idioma: En Ano de publicação: 2024 Tipo de documento: Article
Texto completo
Imprimir
XML
PubMed Links
Buscar no Google

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: RNA / Núcleo Celular Limite: Animals / Humans Idioma: En Ano de publicação: 2024 Tipo de documento: Article