10x genomics chromium gem-x single cell 3' v4 gene expression with feature barcoding  (10X Genomics)

Journal: bioRxiv

Article Title: Single-Nucleus Neuronal Transcriptional Profiling of Male C. elegans Uncovers Regulators of Sex-Specific and Sex-Shared Behaviors

doi: 10.1101/2024.12.12.628226

Figure Lengend Snippet: (A) Representative image of pan-neuronal GFP tagged him-8 males ( Prgef-1::his-58::GFP;him-8) . (B) 6-well tissue culture inserts with modified filters were used to separate sexes in C. elegans, achieving an average ∼95% purity for both male and hermaphrodite fractions. (C) Overview of nuclei isolation and sequencing. Pan-neuronal histone GFP-tagged him-8 males and hermaphrodites were separated, nuclei were isolated, FACS sorted for Hoescht and DAPI positive nuclei, followed by barcoding, cDNA amplification, and library preparation using 10X genomics chromium X, then Illumina RNA sequenced. (D) UMAP of 61,936 male-specific neurons that passed quality control. (E) Feature plot of pkd-2, clec-164, flp-17, and Y70G10A.2 expression (known markers expressed in CEM, HOB, and ray neurons).

Article Snippet: Single nuclei suspension samples were then provided to the Princeton Genomics Core for 10X Genomics Chromium X barcoding, cDNA amplification, library preparation and Illumina next generation sequencing, as described in St. Ange and Weng et al. 2024 .

Techniques: Modification, Isolation, Sequencing, Amplification, Control, Expressing