GSM8184621: Human PBMCs_2; Homo sapiens; RNA-Seq
1μg/ml CD3 and CD28 were used to activate Human PBMCs after thawing them. The control group was added to PBS for 4h, and the experimental group was treated with the same amount of ITPRIPL1-ECD-Fc for 4h. Then prepared single-cell suspensions and measured cell viability. Gel beads containing Barcode information are mixed with a mixture of samples and enzymes using the 10x Genomics and then coated with droplets of oil surfactants located in the Chromium system to form GEMs(Gel Beads in-Emulsions). In GEMs, Gel beads will release primers containing Illumina TruSeq Read1, 10x Barcode, UMI and Poly(dT), and bind with mRNA released after cell lysis to form full-length cDNA with Barcode label through reverse transcription. cDNA in GEMs was recovered and amplified by PCR. Then the cDNA fragment length was optimized by enzyme digestion and fragment screening. Finally, the construction process was completed through terminal repair and the traditional second-generation sequencing connector, and the standard sequencing DNA library was amplified by PCR.