Single-Cell RNA Sequencing Protocol

Single-cell suspensions (1 × 10⁵ cells/mL) in PBS were loaded into microfluidic devices using the Singleron Matrix^® Single Cell Processing System (Singleron, China). Subsequently, the scRNA-seq libraries were constructed according to the protocol of the GEXSCOPE^® Single Cell RNA Library Kits (Singleron Biotechnologies) [25 (link)]. Individual libraries were diluted to 4 nM and pooled for sequencing. Finally, pools were sequenced on Illumina NovaSeq 6000 (Illumina, San Diego, CA, USA) with 150-bp paired end reads.

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Du J., Cheng N., Deng Y., Xiang P., Liang J., Zhang Z., Hei Z, & Li X. (2023). Astrocyte senescence-like response related to peripheral nerve injury-induced neuropathic pain. Cellular & Molecular Biology Letters, 28, 65.

Publication 2023

Matrix® Single Cell Processing System GEXSCOPE® Single Cell RNA Library Kits NovaSeq 6000

Cell Library Microfluidic devices Scrna seq

Corresponding Organization :

Other organizations : Third Affiliated Hospital of Sun Yat-sen University, Sun Yat-sen University, Southern Medical University, Nanfang Hospital

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Variable analysis

independent variables

Microfluidic devices used for cell processing
GEXSCOPE® Single Cell RNA Library Kits used for library construction
Sequencing platform (Illumina NovaSeq 6000)

dependent variables

Single-cell RNA sequencing (scRNA-seq) libraries

control variables

Cell suspension concentration (1 × 10^5 cells/mL) in PBS
Sequencing read length (150-bp paired end)

controls

No positive or negative controls were explicitly mentioned in the input protocol.

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