Single-cell RNA-seq of alpha-synuclein-expressing human H4 cells
scRNA-seq libraries were prepared according to the Chromium Single Cell 3 Reagent Kits v3.1 with Feature Barcoding Technology User Guide (CG000389, 10x Genomics). For the generation of the barcode specific library the MULTI-seq additive primer (5-CCTTGGCACCCGAGAATTCC-3) was added during cDNA amplification. After which, barcode and endogenous cDNA fractions were separated using SPRI size selection, allowing for the construction of an independent MULTI-seq barcode and a gene expression library. For the endogenous transcript fraction, the protocol was continued according to the user guide. For the MULTI-seq barcode fraction, an additional 2x SPRI clean-up was performed. Lastly, an indexing PCR was performed to add Illumina adapters and sample specific indexes using the Feature SI Primer 3 (10x Genomics, PN2000263) and TruSeq RPIX primer. After library quality control using a Bioanalyzer high sensitivity DNA analysis kit (Agilent, 5067-4626), libraries were sequenced on an Illumina NovaSeq6000 and an Illumina NextSeq2000 instrument, with 28 and 91 base-pairs for read 1 and 2, respectively. The FASTQ files provided here contain only the barcodes used in the analysis after filtering.
SUB16360043
asyn_S1