title: GSM6696870: PAMs, Mock replicate 1; Sus scrofa; RNA-Seq
library_name: GSM6696870
library_construction_protocol: For in vivo study, PAM cell were collected directly from infected or control pigs by lung lavage. For in vitro study, cells were collected from culture. All cells were washed, and resuspended in 1X PBS to a final concentration of about 1,200 cell/uL before performing library construction. Library was performed according to the manufacter's instructions (single cell 3' v3 protocol, 10x Genomics). Briefly, GCs 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.