GSM8321128: OVE26 Lrg1-/- 1; Mus musculus; RNA-Seq
To optimize the representation of all kidney cells, glomeruli were first isolated from harvested mouse kidneys using magnetic beads (16), and kidney cells from glomerular and non-glomerular fractions were prepared separately and pooled together per mouse kidney prior to downstream processing. The glomerular fraction was prepared as described previously, and the non-glomerular fraction was prepared with the following modifications: harvested fraction was incubated in the digestion buffer for 10 min at 37°C, and dissociated cells were filtered through 40-micron cell strainers. Kidney single-cells were processed using the Chromium Single Cell 3' kit (v3) of 10x Genomics according to the manufacturer's protocol. The final constructed single-cell libraries were sequenced by Illumina Novaseq machine with total reads per cell targeted for a minimum of 30,000.