Single-Cell RNA Sequencing Library Preparation

Single-cell suspensions (1×10⁵ cells/ml) with PBS (HyClone) were loaded into microfluidic devices using the Singleron Matrix^® Single Cell Processing System (Singleron). Subsequently, the scRNA-seq libraries were constructed according to the protocol of the GEXSCOPE^® Single Cell RNA Library Kits (Singleron) 14 (link). Individual libraries were diluted to 4 nM and pooled for sequencing. At last, pools were sequenced on Illumina HiSeq X with 150 bp paired end reads.

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Li X., Zhao S., Bian X., Zhang L., Lu L., Pei S., Dong L., Shi W., Huang L., Zhang X., Chen M., Chen X, & Yin M. (2022). Signatures of EMT, immunosuppression, and inflammation in primary and recurrent human cutaneous squamous cell carcinoma at single-cell resolution. Theranostics, 12(17), 7532-7549.

Publication 2022

Singleron Matrix® Single Cell Processing System GEXSCOPE® Single Cell RNA Library Kits HiSeq X

Cell Library Microfluidic devices Scrna seq

Corresponding Organization :

Other organizations : Hunan Cancer Hospital, Xiangya Hospital Central South University, Central South University

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Variable analysis

independent variables

Singleron Matrix® Single Cell Processing System

dependent variables

ScRNA-seq library construction
Sequencing (Illumina HiSeq X, 150 bp paired-end reads)

control variables

PBS (HyClone) used for cell suspensions
Cell concentration (1×10^5 cells/ml)
GEXSCOPE® Single Cell RNA Library Kits (Singleron) used for library construction
Library dilution to 4 nM and pooling for sequencing

controls

No positive or negative controls explicitly mentioned

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