GSM8003491: C4; Mus musculus; RNA-Seq
Total cells were sorted from MedLNs of C4-treated or -untreated OVA-induced asthma model. The single cells were obtained by the method of grinding through a 70 μm cell strainer. For the quality check and counting of single cell suspension, the cell survival rate is generally above 80%. The cells that have passed the test are washed and resuspended to prepare a suitable cell concentration of 700~1200 cells/ul for 10x Genomics ChromiumTM. Library construction was performed according to the manufacture's protocol(Single Cell 3′ library and Gel Bead Kit v.3).Briefly, GEMs (Gel Bead in Emulsion) were constructed for single cell separation according to the number of cells to be harvested. After GEMs were normally formed, GEMs were collected for reverse transcription in a PCR machine for labeling. The GEMs were oil-treated, and the amplified cDNA was purified by magnetic beads, and then subjected to cDNA amplification and quality inspection. The 3ʹ Gene Expression Library was constructed with the quality-qualified cDNA. After fragmentation,  adaptor ligation, sample index PCR, etc., the library is finally quantitatively examined.