GSM8783538: dKO; Mus musculus; RNA-Seq
This study was involved samples pooled together for single cell sequencing. We  pooled 3 samples together as control (wild type or heterozygote), and 4 samples  as dKO (Emx1-Cre Mcmbp/Trp53 double conditional KO). Single-cell RNA processing was performed using SeekOne® Digital Droplet (SeekOne® DD) system and associated kits from SeekOne Bio, including: SeekOne® DD S3 Microarray Kit (SEEKGENE, K00202-0201), SeekOne® DD Single-Cell Purification Kit (SEEKGENE, K00202-0205), SeekOne® DD Single-Cell Whole-Sequence Microbead Kit (SEEKGENE, K00801-0202), SeekOne® DD Single-Cell Full Sequence Reverse Transcription Kit (SEEKGENE, K00801-0203) and SeekOne® DD Library Construction Kit (SEEKGENE, K00202-0204). The SeekOne® DD platform enables comprehensive processing from single-cell nucleic acid labeling to full-sequence transcriptome library construction. The system employs microfluidic-based water-in-oil droplet technology for single-cell isolation and capture. RNA molecules from individual cells are labeled using nucleic acid-modified Barcoded Beads, and full-length coding and non-coding RNAs are captured using random primers. Intracellular rRNA and mitochondrial contamination are eliminated using proprietary blocker technology. Libraries were constructed following the "SeekOne® DD Single-Cell Whole Sequence Transcriptome Kit Instruction Manual" to ensure compatibility with high-throughput sequencers. Quality control criteria for qualified libraries included: main peak fragment size range: 450-900 bp, absence of small fragments (verified by Agilent 4200 TapeStation), Minimum library concentration: 1 ng/μL (measured by Qubit 4.0). If small fragments were detected, additional 0.6x purification steps were performed until complete removal was achieved.