GSM6784569: Sample#3 (Up Epith esophagus); Mus musculus; RNA-Seq
Tissues were digested and sorted in 1ml PBS 0.04% BSA. Samples were loaded on a chromium chip and processed using the Chromium Single Cell 3' Reagent kits v3 (10x Genomics) following manufacturer's recommendations. Multiplexed libraries were obtained following manufacturer's recommendations of Chromium Single Cell 3' Reagent kits v3 (10x Genomics) and loaded on a Novaseq 6000 (Illumina). Briefly, samples 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.