Benchmarking Single-Cell mRNA-Sequencing Technologies Uncovers Differences in Sensitivity and Reproducibility in Cell Types With Low RNA Content.
J Biomol Tech
; 32(4)2021 12 15.
Article
en En
| MEDLINE
| ID: mdl-35837268
Single-cell RNA sequencing (scRNA-seq) has the ability to classify each cell and determine the transcriptomic profile of specific cell types and cells of a given disease state; however, sensitivity of the gene count for each cell can be a critical component to the success of a single-cell study. The recently introduced SMART-Seq Single Cell PLUS Kit (SSsc PLUS) claims to provide higher sensitivity and reproducibility versus popular methods for the sequencing analysis of single cells. Here, the cDNA-generation component of the kit, SMART-Seq Single Cell Kit (SSsc), was compared with the popular homebrew protocol, Smart-seq2, and its update, Smart-seq3. The SMART-Seq Library Prep Kit from SSsc PLUS was benchmarked against a commonly used scRNA-seq library preparation method, Illumina Nextera XT. Finally, the SSsc chemistry was tested in both full and fractional volumes on 2 popular liquid-handler devices to investigate whether the high sensitivity was maintained in miniaturization. We demonstrate that SSsc PLUS outperforms these other full-length methods in convenience, sensitivity, gene identification, and reproducibility while also offering full compatibility with automation platforms.
Palabras clave
Texto completo:
1
Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
ARN
/
Análisis de la Célula Individual
Tipo de estudio:
Diagnostic_studies
/
Prognostic_studies
Idioma:
En
Revista:
J Biomol Tech
Asunto de la revista:
BIOTECNOLOGIA
Año:
2021
Tipo del documento:
Article
País de afiliación:
Estados Unidos
Pais de publicación:
Estados Unidos