Deciphering the Single-Cell Systemic Immune Landscape in Oral Lichen Planus

Single Cell 3' libraries were prepared using standard Illumina paired-end constructs, which include the P5 and P7 adaptors. The libraries incorporated a 16 bp 10x Barcode and a 12 bp Unique Molecular Identifier (UMI) within Read 1. The cDNA fragments in the 3' Gene Expression libraries were sequenced using Read 2, while the DNA from Cell Multiplexing Oligo Feature Barcodes in the Cell Multiplexing libraries was sequenced using Read 2N. The libraries were subjected to paired-end sequencing on an Illumina platform. Sample indexing was achieved using i7 and i5 index sequences. The following standard Illumina sequencing primer sites were utilized: for the 3' Gene Expression libraries, TruSeq Read 1 and TruSeq Read 2 primers were employed; for the Cell Multiplexing libraries, Nextera Read 1 and Nextera Read 2 primers were used. This method ensured accurate and efficient sequencing of both gene expression and multiplexing features, facilitating comprehensive single-cell analysis.