GSM9448800: FACS-purified MG and their neuronal progeny 1 month after NMDA and Adezmapimod (MAPK14 inhibitor) treatment; Mus musculus; RNA-Seq
Cell quality control adjust the cell concentration to the optimal level, typically 1000 cells/μL. 10X Barcoding of cDNA Fragments Gel beads carrying barcode information are combined with a mixture of cells and enzymes, then encapsulated by oil-surfactant droplets within a microfluidic system to form GEMs. The GEMs flow into a collection reservoir, where the gel beads dissolve to release the barcode sequences. Reverse transcription is performed to generate cDNA fragments, and samples are labeled. The gel beads are then disrupted and the oil droplets are broken. Using the cDNA as a template, PCR amplification is carried out. The products from all GEMs are pooled to construct a standard sequencing library. The cDNA is enzymatically fragmented into segments of approximately 200–300 bp. This is followed by traditional second-generation sequencing library preparation steps, such as the addition of sequencing adapters and primers. Finally, PCR amplification is performed to obtain the DNA library.