GSM9336036: mRNA_HFD6w_2; Mus musculus; RNA-Seq
ApoE knockout mice subjected to either high-fat diet alone or high-fat diet combined with chronic restraint stress for 6 weeks underwent spleen homogenisation to obtain a single-cell suspension. Single-cell suspension of spleen was obtained and analyzed using scRNA-seq. Cell Quality Control: Adjust the cell concentration to the desired level, typically 700–1200 cells/μL. Digest cDNA into fragments of approximately 200–300 bp, incorporate sequencing adapters and primers, and perform conventional second-generation sequencing library preparation. Finally, conduct PCR amplification to obtain the DNA library. Immune repertoire library construction: TCR/BCR constant regions are enriched via nested PCR using nested primers. Fragmentation is then performed with controlled cleavage lengths to generate sequences covering the entire V(D)J fragment cDNA. Through end-repair, A-adding, and adapter ligation with Read2 sequencing primers, a V(D)J library containing P5 and P7 adapters is constructed.