GSM8685226: Nuclei from P3 pooled from mouse kidneys; Mus musculus; RNA-Seq
Kidneys were dissected in cold PBS without calcium or magnesium, snap-frozen for genotyping, and then thawed on ice for pooling. We minced the kidneys using razorblades and homogenized them with a Dounce homogenizer in Nuclei EZ Lysis buffer (Sigma) supplemented with protease inhibitor (Roche), RNase inhibitor (Promega and Life Technologies). The homogenate was filtered through a 40μm cell strainer (pluriSelect) and centrifuged at 500 x g for 5 min at 4°C. The pellet was resuspended, washed, and filtered again through a 5μm cell strainer (pluriSelect). The resulting nuclei suspensions were submitted to UTSW Next Gen Sequencing Core for library preparation and sequencing using the Chromium Single Cell 3' Prime Gene Expression kit (10x Genomics) on an Illumina NextSeq 2000.