GSM7050665: Control, replicate 1, scRNAseq; Mus musculus; RNA-Seq
On postnatal day 14, the ligated cerebral cortex of the experimental group and the ipsilateral cerebral cortex of the control group were collected (1) Nuclear quality inspection After resuspension of the cell nuclei, the cells were collected by filtration through a 20um cell sieve and counted. (2) 10X labeled cDNA fragments Gel Beads containing barcode information are bound to a mixture of cells and enzymes and then coated with oil surfactant droplets located In a microfluidic "double cross" system to form GEMs (gel beads-in-emulsions). GEMs flow into the reservoir and are collected, gel beads dissolve to release Barcode sequences, reverse transcribe cDNA fragments, and label the samples. Gel beads were broken up and oil droplets were broken up, and cDNA was used as template for PCR amplification. The products of all GEMs were pooled to construct a standard sequencing library. (3) Construction of standard sequencing library First, the cDNA fragment was broken into 200-300 bp fragments by enzyme digestion, and the library construction process of traditional second generation sequencing was added, such as sequencing adapter and primer. Finally, the DNA library was amplified by PCR. (4) Library sequencing High-throughput sequencing was performed on the constructed library using the double-end sequencing mode of the Illumina sequencing platform.