title: GSM6733477: Cγ1ATG7fl/fl  (Exp), 10d.p.i, Replicate 1; Mus musculus; RNA-Seq
library_name: GSM6733477
library_construction_protocol: Freshly collected total splenic cells were isolated from spleens of 10- or 21-days immunized mice by tissue homogenization. Cells were resuspended in staining buffer (5% FBS in PBS 1X) and preserved at 4C during all process. Red blood cells were lysed using RBC lysis buffer (Biolegend). Library was performed according to the manufacter's instructions (Chromium Next GEM Single Cell 3'Kit v3.1, 10x Genomics). Briefly, splenocytes were resuspended in the master mix and loaded together with partitioning oil and gel beads into the chip to generate the gel bead-in-emulsion (GEM). The poly-A RNA from the cell lysate contained in every single GEM was retrotranscripted to cDNA, which contains an Ilumina R1 primer sequence, Unique Molecular Identifier (UMI) and the 10x Barcode. The pooled barcoded cDNA was then cleaned up with Silane DynaBeads, amplified by PCR and the apropiated sized fragments were selected with SPRIselect reagent for subsequent library construction. During the library construction Ilumina R2 primer sequence, paired-end constructs with P5 and P7 sequences and a sample index were added.