title: GSM8951844: Sptbn5_ko_heart; Mus musculus; RNA-Seq
library_construction_protocol: E10.5 wild-type and homozygous ciliary gene knockout/mutant mouse embryos were isolated and dissected for cardiac OFTs. The obtained tissues were washed for 2-3 times with DPBS pre-cooled at 4℃ and then was transferred to a 2 mL centrifuge tube with tissue storage buffer (Miltenyi Biotec, #130-100-008). The tube was centrifuged at 50 × g for 1 min at 4 °C, after which the tissue storage buffer was discarded and 1 ml of pre-warmed digestion solution (Miltenyi Biotec, #130-110-203; 37 °C) was added to the OFT tissue. The tissues were incubated in a 37 °C water bath and gently pipetted every 5 min, for a total digestion time of approximately 15 min. Enzymatic digestion was terminated by adding 10% FBS (Pricella, #164210-50), and the cell suspension was filtered through a 40 µm cell strainer (Corning, #CLS431750). The filtered suspension was centrifuged at 300 × g for 3 min at 4 °C, and the pellets were resuspended in PBS containing 0.01% Bovine Serum Albumin (Sigma, #A1933). Libraries were constructed using the High Throughput Single Cell 3'RNA-Seq Kit v2.0 (MobiDrop, #S050200201) and indexed using the 3' Single Index Kit (MobiDrop, #S050300201).