GSM9759673: Normal; Homo sapiens; RNA-Seq
The isolated endometrial cells were resuspended in digestion buffer  and incubated at 37 °C for 30 min. Cells were washed, re-suspended in PBS, and stained with a viability dye. After washing, cells were resuspended in FACS buffer . Live endometrial cells were sorted using a FACS Jazz (Becton Dickinson) based on the presence or absence of mCherry signal, using a C57BL/6 mouse as a negative control. Library was performed according to the manufacter's instructions (single cell 3' v2 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 llumina 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 llumina R2 primer sequence, paired-end constructs with P5 and P7 sequences and a sample index were added.