GSM8666152: C80 scRNA-seq of WT mouse lung; Mus musculus; RNA-Seq
The entire lung tissues were harvested from euthanized mice, immediately placed in pre-chilled RPMI 1640 medium for initial washing to remove blood residues. The tissues were washed twice with PBS, cut into 1-2mm³ pieces, and enzymatically digested using SeekGene Tissue Dissociation Reagent A, Collagenase IV (0.1 mg/ml), and DNase I (0.01 mg/ml) in a 37°C water bath for 30 minutes to dissociate lung tissues into single-cell suspensions. Single-cell RNA libraries were prepared using the SeekOne® Digital Droplet Single Cell Library Preparation Kit (SeekGene K00202). Cells were mixed with reverse transcription reagents and loaded into SeekOne® chips, followed by the addition of barcoded hydrogel beads and separation oil. After droplet generation, reverse transcription was performed at 42°C for 90 minutes, and PCR amplification was carried out to prepare the sequencing library.